Newswise In August of 2017, SPARGO, Inc. will assume responsibility for the management of exhibit and sponsorship sales for the American Society for Cell Biologys 2017 and 2018 ASCB|EMBO Meetings and 2019 and 2020 ASCB Annual Meetings. Additionally, SPARGO will sell advertising in ASCBs three publications and other digital communications.

The Annual Meeting, which has seen steady attendance growth over the past four years, brings together scientists from all over the world to discuss new experimental results and techniques in various domains of basic science and creates the environment for broader discussions on topics ranging from what is cell biology to the future of biomedical research, funding, training, and publishing. The Learning Center at the Annual Meeting will continue to feature technical and scientific exhibits, integrate the poster sessions, and host the popular Tech Talks and microsymposia presentation theaters in which cutting edge content is delivered by the exhibiting companies.

The 2017 ASCB|EMBO meeting will be held December 2-6 in Philadelphia, PA. The Northeast/Mid-Atlantic region boasts a large concentration of medical and academic institutions. Holding the conference in this region makes it affordable and convenient for both domestic and international scientists to attend.

We are proud to partner with SPARGO, says Alison Harris, ASCB's Director of Meetings. Our aim is to continually improve the meeting experience for attendees and exhibitors, and were confident that SPARGOs experienced team of event management professionals will help us fulfill this mission.

We are honored to have been selected as a partner to ASCB. We will focus on exhibitor satisfaction, improved ROI, and show growth and capitalize on the advertising opportunities as a key strategy for exhibitors to have a year-long exposure to the influential members of the ASCB community, says Susan Bracken, President and CEO at SPARGO, Inc.

About ASCB:

The American Society for Cell Biology (ASCB) was founded in 1960 to bring the varied facets of cell biology together. The Society's purpose is to promote and develop the field of cell biology. ASCBs mission is to: advance scientific discovery, advocate for sound research policies, improve education, promote professional development, and increase diversity in the scientific workforce. Its objectives are achieved through the scholarly dissemination of research at its Annual Meeting and in its publications. See more at http://www.ascb.org.

About SPARGO, Inc.:

SPARGO is a full-service event management company. SPARGO offers a full suite of services that support the production of tradeshows, conventions, conferences, symposiums, and seminars. See more atwww.spargoinc.com.

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The American Society for Cell Biology (ASCB) Appoints SPARGO, Inc. as Official Exposition and Advertising Sales and ... - Newswise (press release)

The Royal Microscopical Society has announced changes to its governing Executive Committee following a recent AGM.

Dr Peter OToole from the University of York has been appointed as Vice-President of the Society and Professor Maddy Parsons has become the new Honorary Secretary for Biological Science.

As Vice-President, Dr OToole will be supporting the current President Professor Michelle Peckham in forming the strategy for the RMS and managing its ongoing activities. With over 1500 members worldwide and activities including an annual calendar of around 20 conferences and training courses, a scientific journal and an active Primary School scheme reaching thousands of children each year, this is a big responsibility to undertake.

Dr Peter O'Toole heads the Imaging and Cytometry Labs within the Technology Facility at the University of York which includes an array of confocal microscopes, flow cytometers and electron microscopes. His research is currently focused on both technology and method development of novel probes and imaging modalities. He has ongoing collaborations with many leading microscopy and cytometry companies and his group also provides research support to many academics and commercial organisations. Dr OToole is also heavily involved with teaching microscopy and flow cytometry which includes organising and teaching on both the RMS Light Microscopy Summer School and the RMS Practical Flow Cytometry courses.

Professor Michelle Peckham, President of the RMS said Im really pleased that the Society has elected Dr OToole as the new Vice-President. Dr OToole has done an excellent job of co-chairing the mmc-series as well as emc2012 when we hosted the European Congress in Manchester. I believe that his passion for teaching and his work as an advocate for Imaging Facility Managers will make him a really great leader for the Society and I look forward to working with him further over the next 3 years.

As Honorary Secretary for Biological Science, Professor Parsons will be championing life sciences in the activities of the RMS as well as acting as Life Sciences Chair for their successful flagship mmc-series, the next event being mmc2019.

Professor Parsons is Professor of Cell Biology at Kings College London. Maddy was awarded a Royal Society University Research Fellowship in 2005 to establish her own group within the Randall Division of Cell and Molecular Biophysics at Kings College London. Professor Parsons has established collaborations with developmental biologists and clinical researchers to study adhesion receptor signalling in skin blistering, wound healing, inflammation and cancer. She works closely with physicists, biophysicists and other world-leading cell migration groups in the field to develop and apply new imaging technologies to dissect spatiotemporal cytoskeletal signalling events in live cells, tissues and whole organisms. As a result of her interest and applications of advanced microscopy, Professor Parsons developed a strong working partnership with Nikon, which subsequently led to the establishment of the state-of-the-art, world-class Nikon Imaging Centre at Kings College London of which she is Director. Professor Parsons also currently works alongside other biotech and pharmaceutical companies to develop and apply advanced imaging approaches to basic mechanisms that underpin drug discovery.

Allison Winton, Chief Executive of the RMS said Professor Parsons has been an active member of the RMS Life Sciences committee for many years and has helped to organise both our Frontiers in BioImaging and Abercrombie meetings identifying key emerging topics and developing valuable links between the Society and many eminent scientists working in the field. Professor Parsons proactive and approachable nature as well as her vast knowledge and experience in the field will make her a great representative for biological science in the Society.

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New Executive Committee Members at RMS - Labmate Online

Almost 54,000 new cases of breast cancer were diagnosed in the UK during 2013, according to Cancer Research UK figures.

In 2014, 11,400 people died from the disease.

Now scientists are hoping more lives could be saved following a medical breakthrough.

Scientists part-funded by Breast Cancer Now, the Medical Research Council (MRC) and other collaborators have developed an innovative imaging technique that could predict whether breast cancer will spread to the lung.

In a new study published in Theranostics, researchers have demonstrated in mice that a new non-invasive imaging method can be used to detect changes in the lungs that signal breast cancer may soon spread there before any metastases are visible.

If given the green-light for use in humans, this approach could enable patients to be offered more intensive therapy earlier, to potentially prevent breast cancer spread.

Dr Fabian Flores-Borja, Research Fellow at the Breast Cancer Now Research Unit at Kings College London said: By combining cell biology and imaging techniques, we have established a method to predict, at an early time-point during tumour development, whether tumour invasion will occur.

We envision this technique being used to help select patients for either further surveillance or intensified therapy, as well as aiding cancer research.

The development of a test that is able to identify an increased risk of metastasis soon after a patient is diagnosed with breast cancer, would be very useful in helping choose the best treatment for patients.

Previous research has shown that the gathering of a special type of immune cell called myeloid-derived suppressor cells (MDSCs) in locations such as the lung prepares the ground for breast cancer metastasis - spread.

This is because the local immune system is suppressed promotes the formation of new blood vessels - a condition called angiogenesis.

WHAT ARE THE SYMPTOMS OF BREAST CANCER?

Researchers at Kings College London have now developed a radioactive tracer molecule to detect MDSCs accumulating in the lung in preparation for the arrival of breast cancer cells and the formation of metastases.

Baroness Delyth Morgan, Chief Executive at Breast Cancer Now, has hailed the news as incredibly exciting.

While more research is needed before this could be tested in patients, the prospect of a hospital scan which could predict whether breast cancer will spread to the lungs is incredibly exciting.

More immediately, this study brings a brand new method to the table that will help researchers unpick how the immune system is involved in the spread of breast cancer.

Finding ways to predict and halt the spread of the disease will be crucial if we are to finally stop people dying from it.

This is a promising step towards being able to use 3D imaging to help offer more personalised therapy. Ultimately, anything that could provide patients and their doctors with a more accurate picture of whether their breast cancer may spread will help us tailor treatments to stop this from happening.

Dr Mariana Delfino-Machin, MRC Programme Manager for Cancer, said the research paves the way for new treatment in the clinic.

"Innovative, non-invasive imaging methods like this, which can help predict and diagnose disease as early as possible and avoid the discomfort of current invasive tests, have the potential to greatly impact cancer treatment and outcomes, she said.

Experts said more studies are now required to develop a more effective tracer molecule - suited for use in humans - to be tested in future clinical trials.

SIX BREAST CANCER MYTHS BUSTED

Read the rest here:
Breast cancer breakthrough: This technique could predict if disease will spread to lungs - Express.co.uk

The researchers discovered that the genetic activity of mouse skeletal muscles is particularly intense during the first two weeks after birth; a number of genes alter the amount of proteins produced, while other genes go through alternative splicing and produce different proteins.

Among the genes going through alternative splicing, those involved in calcium-handling functions predominated. Calcium is very important for skeletal and heart muscle because the influx of calcium into the cell stimulates contraction and other functions.

First author Dr. Amy Brinegar, who was a graduate student in the Cooper lab while she was working on this project and recently graduated from the doctoral program in molecular and cellular biology at Baylor, selected three calcineurin A genes, which are involved in calcium-handling functions, and reversed their natural process of alternative splicing in adult mouse muscles. Then, Dr. George Rodney, associate professor of molecular physiology at Baylor, and a graduate student in his lab, James Loehr, who are co-authors on this paper, determined the effect of switching back alternative splicing on functions of isolated adult mouse skeletal muscle in the lab.

They discovered that muscles in which the adult forms of the calcineurin A genes had been switched back to the newborn forms showed a change in calcium flow and were less strong than muscles that retained the adult forms of calcineurin A.

We showed that just by changing three of about 11,000 genes that are estimated to be expressed in adult mouse muscle, we were able to change physiological parameters of those muscles, said Brinegar. This work supports the growing evidence in favor of a physiological role of alternative splicing.

Importantly, about 50 percent of the genes we discovered to undergo alternative splicing are conserved, meaning that the genes go through the same changes both in mice and humans, which opens the possibility of modeling human muscle disorders in the mouse, Cooper said.

Other contributors top this work include Zheng Xia and Wei Li, both from Baylor.

Financial support was provided by National Institutes of Health grants R01AR045653, R01HL045565, R01AR060733, T32 HL007676, R01HG007538, R01CA193466 and R01AR061370. Further support was provided by the Muscular Dystrophy Association grant RG4205.

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Change in protein production essential to muscle function - Baylor College of Medicine News (press release)

Maui High School graduate Phyllis Raquinio stands by her research project on the connection between Type 2 diabetes and cancer survival rates at the University of Hawaii Cancer Center. Raquinio was one of 19 interns selected to conduct research at the center over the summer. Raquinio majors in molecular cell biology and will be a senior at UH-Manoa in the fall. University of Hawaii Cancer Center photo

Instead of enjoying a lazy, carefree vacation, Maui High graduate Phyllis Raquinio was hard at work this summer poring over cancer data, discussing how hula and physical activity can help post-survival cancer patients and learning how advertisements can play a role in the growing consumption of e-cigarettes.

Raquinio was one of 19 students selected and the only Maui graduate to conduct research as an intern at the University of Hawaii Cancer Center.

I did not know a lot about cancer and wanted to learn more about it to have a better idea on how cancer research is conducted, Raquinio said via email Sunday. I was surprised to be accepted in the internship, considering how competitive it was, but I was extremely excited to start working and helping expand my knowledge of cancer.

The UH Cancer Center is one of 69 research institutions designated by the National Cancer Institute. It started an internship program in 2004, said Yvette Amshoff, education and outreach coordinator at the Cancer Center.

While many of (the interns) are interested in careers as doctors, we aim to show the many opportunities in cancer research and how research is important to enforcing new regulations, clinical practices and cancer cures in the health setting, Amshoff explained.

According to the center, interns are chosen from public and private schools from across the state and the nation in a highly competitive process. This year, 19 out of 74 applicants were selected, with an average grade point average of 3.77. Program funding comes from the National Cancer Institute, the Meiji Yasuda Life Insurance Co. and the Friends of the Cancer Center.

Interns work under the guidance of faculty members and learn about new advances in research and technology. At the end of the program, each intern submits a project on a topic of his or her choice. Joe Ramos Jr., deputy director and professor at the center, explained that students can either explore cancer biology, which takes a look at the molecular mechanisms behind cancer, or population sciences in the Pacific.

Students like Ms. Raquinio who go into the population sciences in the Pacific projects are examining how the environment (what we eat, smoke or are otherwise exposed to in Hawaii) and our genes and behaviors interact to affect cancer risk, cancer progression and survival outcomes, Ramos said.

For example, Ms. Raquinio examined how having type 2 diabetes affected survival of patients with breast cancer and colon cancer. This is part of a continuing project on this topic. If we can understand these connections better, we can better prevent people in Hawaii and the Pacific from getting cancer or improve their survival chances.

Born and raised on Maui, Raquinio graduated from Maui High School in 2014. She said a few of her friends interned at the center last summer and encouraged her to apply. While her grandfather and a close instructor suffered from cancer, Raquinio said her interest stemmed mainly from the biology behind cancer. At UH, she majors in molecular cell biology with a minor in English.

Over the summer, Raquinio worked with Dr. Gertraud Maskarinec, analyzing data from a large, multiethnic population study in Hawaii. The study included more than 215,000 participants. Of that group, 5,000 had been diagnosed with colorectal cancer and 7,500 had been diagnosed with breast cancer.

Phyllis was a very dedicated intern, Amshoff said. Learning how to use statistical software to analyze a large data set can be very tricky, but Phyllis caught on quickly, and now her findings are being refined to be submitted to a journal.

Raquinio said the study produced unexpected results but explained that she was still completing her paper and working to better understand the results before reporting them.

As an intern, Raquinio said she learned a lot from interns researching other topics and gained a better understanding of the process for clinical studies and why it takes a long time for treatments to be approved.

This internship has opened my mind to potentially pursuing cancer research, Raquinio said. It has helped me understand more about specific cancers, how biology plays a role, and the processes to conducting research for treating cancer.

Raquinio said she hopes to enroll in medical school after she graduates, with eventual plans to become a doctor.

Ms. Raquinio and the other students bring a fresh perspective and enthusiasm to the projects that makes working with them tremendously rewarding for the mentors, Ramos said. I am consistently impressed that these students come in with very little to no experience working in cancer research and quickly get their bearings and leave with a real appreciation for the kind of disciplined inquiry and experimentation required to make important advances in understanding and treating cancer.

* Colleen Uechi can be reached at cuechi@mauinews.com.

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Maui High graduate spends summer turning raw data into cancer research - Maui News

Researchers say they've found a system in the human heart that allows the organ to restart itself. Their discovery could lead to the replacement of pacemakers.

In an episode of Star Trek: The Next Generation, Lt. Worf is badly injured, but recovers when it is discovered that his body holds a lot of redundant parts and organs for example, 23 ribs that allow him to regenerate.

Science fiction?

Not entirely.

A team of researchers at The Ohio State University Wexner Medical Center discovered that the human heart contains its own fail-safe backup battery system to regulate the heartbeat.

Their findings were published in Science Translational Medicine.

If further testing is successful, fewer people might need mechanical pacemakers in the future.

The potential market is big.

More than 200,000 people in the United States have a pacemaker implanted every year.

The research is still preliminary, but scientists hope to turn it into practical use some day.

In the future we want to develop something that practitioners would welcome, Vadim Fedorov, PhD, an associate professor of physiology and cell biology at The Ohio State University College of Medicine, told Healthline.

Fedorov explained that an implanted pacemaker works by replacing the hearts defective natural pacemaker functions.

The sinoatrial (SA) node, or sinus node, is the heart's natural pacemaker. It's a small mass of specialized cells in the top of the right atrium (upper chamber of the heart). It produces the electrical impulses that cause the heart to beat.

The heart is hardwired to maintain consistency. Irregular heartbeat, or arrhythmia, can be due to heart disease or other problems, such as changes in diet or hormones or electrolyte imbalance.

Optical and molecular mapping of the human heart revealed that the SA node is home to multiple pacemakers, specialized cardiomyocytes that generate electrical heartbeat-inducing impulses.

Total cardiac arrest occurs only when all pacemakers and conduction pathways fail.

Too technical?

Think of it as a car battery. One day your car wont start. Turns out the battery is still good, but one of the connector cables is bad.

So you clean or replace the wire and save yourself from major repairs.

The Ohio State teams discovery showed that the human heart battery restarts itself.

To prove their point, the researchers actually restarted hearts that were destined for the trash heap.

Most of them came from people getting new hearts or accident victims whose hearts were not suitable for transplant.

We kept them in a special solution, he said. When we warm them to body temperature, they will beat.

The discovery, while exciting, is not going to change clinical practice in the next 60 days.

But it offers promise.

Dr. John Hummel, FACC, is a cardiologist at The Ohio State University Wexner Medical Center and is director of the electrophysiology research section and professor of cardiovascular medicine.

He told Healthline the study is intriguing.

These findings finally give us insight as to the actual structure and behavior of the natural pacemaker of the human heart, he said. Diagnosing disease of the natural pacemaker is often straightforward, but can also be one of the more challenging diagnoses to make.

Dr. Fedorovs findings will likely allow us to develop new approaches to discriminate disease from normal behavior of the sinus node, and give our patients a definitive diagnosis of health or disease of the hearts natural pacemaker, Hummel explained.

Funding to translation of this bench research to clinic research is the next step, he added.

Dr. Gordon Tomaselli, professor of medicine, cellular and molecular medicine at the Johns Hopkins School of Medicine and past president of the American Heart Association, expressed similar thoughts.

The work by Vadim Fedorovs group is a beautifully done study on explanted [not used for transplant] human hearts, Tomaselli told Healthline.

He called the infrared optical mapping studies with pharmacological interventions demonstrating the functional redundancy and complexity of the sinoatrial node (SAN) the most compelling part of the work.

Being able to view the hearts in three dimensions increases the researchs usefulness.

Tomaselli pointed out that researchers have known for decades from previous work in animals, and in clinical human electrophysiological labs, that SAN is functionally redundant and anatomically complex.

He urged caution.

I do not think this paper will fundamentally change the management of patients with regard to pacemaker implantation, he said. Although around half of pacemakers are implanted for diseases of the sinus node or atrium, they are implanted not to prolong life but instead to relieve symptoms [fatigue, shortness of breath particularly with exercise].

He went on, The more life-threatening problems with electrical conduction in the heart for which we put in pacemakers to prolong life involve the electrical system that connects the top and bottom chamber [called the AV node] and the conduction system in the lower chambers. This paper does not address this problem.

So, for the meantime, a Klingon skeleton might be your best bet.

See more here:
The Human Heart May Have a Natural 'Backup Battery' - Healthline

Lysosomes play an important role in cells: They break down old material so the body can dispose of it. Now, scientists at Yale University are zeroing in on lysosomes in the brain, and they believe theyve discovered how these garbage disposal units may contribute to the buildup of the amyloid plaques characteristic of Alzheimers disease.

The Yale researchers discovered defects in the process by which lysosomes travel within neurons. And when this transport goes wrong, lysosomes build up in swollen axons that surround amyloid-beta, the protein thats associated with brain plaques. The team published thefindings in The Journal of Cell Biology.

RELATED: Alzheimer's hopes dashed as Lilly gives up on amyloid drug solanezumab

When lysosomes travel from the ends of axons into the center of neuronal cells, they mature and develop the ability to degrade old cell components. But sometimes they get stuck in swollen axons and fail to mature. The Yale scientists werent sure how this defect contributed to the buildup of amyloid plaque, so they designed an experiment in which they interfered with the transport of lysosomes in mouse neurons.

They discovered that when neurons are deprived of a protein called JIP3, they fail to properly transport lysosomes from the axons, according to a statement. Swollen axons also accumulate amyloid precursor protein (APP) and two enzymes that cause APP to generate amyloid-beta: BACE1 and presenilin 2.

When the team removed a copy of the gene that makes JIP3 from mouse models of Alzheimers, the animals produced more amyloid-beta, and they formed larger plaques surrounded by an increased number of swollen axons.

"Collectively, our results indicate that the axonal accumulations of lysosomes at amyloid plaques are not innocent bystanders but rather are important contributors to APP processing and amyloid plaque growth," said co-author Shawn Ferguson of the Yale School of Medicine in the statement.

The so-called amyloid hypothesis in Alzheimers remains controversial. Many neurological researchers believe amyloid plaques are central players in the disease. But efforts to target those plaques with drugs have been disappointing so far. One of the most high-profile anti-amyloid drugs was solanezumab, Eli Lillys experimental Alzheimers drug, whichthe company abandoned in 2016 after years of failed trials.

But researchers are still investigating new ways of preventing amyloid from building up in the brain. In June, for example, a University of Cambridge team described computer-generated antibodies they developed that prevent amyloid-beta from clumping together and forming plaques.

RELATED: Designer antibodies block Alzheimers plaque from forming

The Yale scientists believe that as they learn more about how deficiencies in lysosome transport contribute to amyloid plaques, they may be able to identify strategies for modulating proteins in the brain to repair the process. Their research might also aid efforts underway to examine how genetics and other risk factorsincluding traumatic brain injuriescontribute to Alzheimers.

Read more:
Cellular 'garbage disposal' units fingered in Alzheimer's development - FierceBiotech

The human brain is routinely described as the most complex object in the known universe. It might therefore seem unlikely that pea-size blobs of brain cells growing in laboratory dishes could be more than fleetingly useful to neuroscientists. Nevertheless, many investigators are now excitedly cultivating these curious biological systems, formally called cerebral organoids and less formally known as mini-brains. With organoids, researchers can run experiments on how living human brains developexperiments that would be impossible (or unthinkable) with the real thing.

Original story reprinted with permission from Quanta Magazine, an editorially independent publication of the Simons Foundation whose mission is to enhance public understanding of science by covering research developments and trends in mathematics and the physical and life sciences.

The cerebral organoids in existence today fall far short of earning the brain label, mini or otherwise. But a trio of recent publications suggests that cerebral-organoid science may be turning a cornerand that the future of such brain studies may depend less on trying to create tiny perfect replicas of whole brains and more on creating highly replicable modules of developing brain parts that can be snapped together like building blocks. Just as interchangeable parts helped make mass production and the Industrial Revolution possible, organoids that have consistent qualities and can be combined as needed may help to speed a revolution in understanding how the human brain develops.

In 2013 Madeline Lancaster, then of the Austrian Academy of Sciences, created the first true cerebral organoids when she discovered that stem cells growing in a supportive gel could form small spherical masses of organized, functioning brain tissue. Veritable colleges of mini-brains were soon thriving under various protocols in laboratories around the world.

Much to the frustration of impatient experimentalists, however, the mini-brains similarity to the real thing only went so far. Their shrunken anatomies were distorted; they lacked blood vessels and layers of tissue; neurons were present but important glial cells that make up the supportive white matter of the brain were often missing.

Worst of all was the organoids inconsistency: They differed too much from one another. According to Arnold Kriegstein, director of the developmental and stem cell biology program at the University of California, San Francisco, it was difficult to get organoids to turn out uniformly even when scientists used the same growth protocol and the same starting materials. And this makes it very difficult to have a properly controlled experiment or to even make valid conclusions, he explained.

Researchers could reduce the troublesome variability by treating early-stage organoids with growth factors that would make them differentiate more consistently as a less varied set of neurons. But that consistency would come at the expense of relevance, because real brain networks are a functional quilt of cell typessome of which arise in place while others migrate from other brain regions.

For example, in the human cortex, about 20 percent of the neuronsthe ones called interneurons, which have inhibitory effectsmigrate there from a center deeper down in the brain called the medial ganglionic eminence (MGE). An oversimplified organoid model for the cortex would be missing all those interneurons and would therefore be useless for studying how the developing brain balances its excitatory and inhibitory signals.

A stained cross section through one of the cortical organoids created by researchers at the Yale Stem Cell Center shows the organization of various cell types into layers of tissue. The organoid is 40 days old in this image. The blue dots are cell nuclei; the red patches are progenitor cells for neurons; the green patches are differentiated neurons.

Courtesy of Yangfei Xiang

Deliverance from those problems may have arrived with recent results from three groups. They point toward the possibility of an almost modular approach to building mini-brains, which involves growing relatively simple organoids representative of different developing brain regions and then allowing them to connect with one another.

The most recent of those results was announced two weeks ago in Cell Stem Cell by a group based at the Yale Stem Cell Center. In the first stage of their experiments, they used human pluripotent stem cells (some derived from blood, others from embryos) to create separate organoid replicas of the cortex and MGE. The researchers then let mixed pairs of the ball-shaped organoids grow side by side. Over several weeks, the pairs of organoids fused. Most important, the Yale team saw that, in keeping with proper brain development, inhibitory interneurons from the MGE organoid migrated into the cortical organoid mass and began to integrate themselves into the neural networks there, exactly as they do in the developing fetal brain.

Earlier this year, teams from the Stanford University School of Medicine and the Austrian Academy of Sciences published reports on similar experiments in which they too developed cortical and MGE organoids and then fused them. The three studies differ significantly in their detailssuch as how the researchers coaxed stem cells to become organoids, how they nurtured the growing organoids, and what tests they ran on the derived cells. But they all found that the fused organoids yielded neural networks with a lifelike mix of excitatory neurons, inhibitory neurons and supporting cells, and that they could be developed more reliably than the older types of mini-brain organoids.

To Kriegstein, all three experiments beautifully illustrate that the cells in organoids will readily transform into mature, healthy tissue if given the opportunity. Once you coax the tissue down a particular developmental trajectory, it actually manages to get there very well on its own with minimal instruction, he said. He believes that specialized organoids could bring a new level of experimental control to neuroscientists explorations: Scientists could probe different brain organoids for information about development within subregions of the brain and then use that combined or fused platform to study how these cells interact once they start migrating and encountering each other.

In-Hyun Park, an associate professor of genetics who led the Yale study, is hopeful that organoids might already be useful in preliminary investigations of the developmental roots of certain neuropsychiatric conditions, such as autism and schizophrenia. Evidence suggests that in these conditions, Park said, there seems to be an imbalance between excitatory and inhibitory neural activity. So those diseases can be studied using the current model that weve developed.

Kriegstein cautions, however, that no one should rush to find clinical significance in organoid experiments. What we really lack is a gold standard of human brain development to calibrate how well these organoids are mimicking the normal condition, he said.

Whatever applications organoid research may eventually find, the essential next steps will consist of learning how to produce organoids that are even more true to life, according to Park. He has also not given up hope that it will eventually be possible to create a mini-brain in the laboratory that is a more complete and accurate stand-in for what grows in our head. Maybe doing so will involve a more complex fusion of organoid subunits, or maybe it will demand a more sophisticated use of growth media and chemicals for directing the organoid through its embryonic stages. There should be an approach to generating a human brain organoid that is composed of forebrain plus midbrain plus hindbrain all together, Park said.

Jordana Cepelewicz contributed reporting to this article.

Original story reprinted with permission from Quanta Magazine, an editorially independent publication of the Simons Foundation whose mission is to enhance public understanding of science by covering research developments and trends in mathematics and the physical and life sciences.

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Lego-Like Brain Balls Could Build a Living Replica of Your Noggin - WIRED

How do initially benign forms of cancer evolve to become aggressive? In a quest to answer this long-standing question, an EU project has studied the growth and clonal evolution of chronic lymphocytic leukaemia (CLL)a blood and bone marrow cancer that mostly starts asymptomatic but can become very aggressive over time.

Cancer evolution is a complex process. Whilst we know that tumour growth is enabled by a continuous process of clonal expansion, genetic diversification and clonal selection, there are still many open questions related to this process. Answering them could be the key to preventing tumour progression and relapses.

According to Dr Michaela Gruber, whose research was funded under the CLL_INCLONEL (Identification and functional dissection of key genetic events in early chronic lymphocytic leukaemia) project, CLL is a valuable model for studying this process due to its high prevalence, initially slow progression and easy access to samples.

Dr Gruber studied the clonal dynamics of a cohort of 21 CLL patients, who were recurrently sampled from diagnosis until the time of first treatment. Her objectives were to identify events leading to disease progression using next-generation sequencing of patient samples. She also developed in vitro models to assess the functional impact of these genetic events on B cell biology, studied their impact on CLL and gathered valuable information on the effects of drugs on potential CLL sub-populations.

Dr Gruber agreed to discuss the project's outcomes and how they could one day lead to individualised diagnostic and therapeutic management of CLL.

What kind of knowledge did you aim to gather from this project?

The key aim of this project was to gain a better understanding of the early dynamics of growth and clonal evolution, as cancer progresses from diagnosis to the need for treatment. CLL is a highly informative model system for studying such natural cancer growth patterns: It typically has a relatively indolent beginning, with potentially long timeframes (in the order of years) before treatment becomes necessary.

Why is it so important to better understand clonal evolution? How can it help prevent tumour progression and relapse?

Insights from recent cancer sequencing studies indicate that the occurrence and expansion of cancer-driving mutations follows a specific sequence. Certain mutations generally appear to occur early in the disease and could be cancer-initiating. Other mutations tend to occur late and appear to have variable impact on tumour expansion. Moreover, different cancer sub-types show different patterns of mutations.

Together, these findings indicate that it could be possible to anticipate the specific evolutionary potential (i.e. plasticity) of a patient's cancer, which actually fuels progression, treatment resistance and relapse. Based on such understanding, therapeutic strategies could be shaped directly against this plasticity of cancer. This would be a major milestone towards overcoming current obstacles to cancer cure.

What would you say were the most important findings from the project?

Our data show that key mutations driving the progression of CLL are established very early in the course of the disease, years before symptoms warrant treatment initiation. For the first time, we were also able to quantify the impact of individual sub-clonal driver mutations on in vivo tumour expansion.

Another important discovery is that of clearly distinguishable growth patterns among patients, both globally as well as on a sub-clonal level. Finally, our data indicate that different patients have different potentials for clonal evolution and growth, and that these patterns remain throughout the entire course of the disease up to the event of relapse.

Can you tell us more about the genome editing technologies you employed?

Suitable experimental models are much needed in order to test the functional impact of observations made in CLL sequencing studies. Thus, we employed novel genome editing strategies, initially using TALENs and then switching to the recently emerged and more easily programmable CRISPR/Cas9 technology. Thanks to the latter, we established an array of isogenic B cell lines, which are used to test the molecular impact of mutations on cellular biology andmost importanttreatment response.

What are your plans now that the project is completed?

We have initiated several follow-up projects in Vienna, which aim to integrate an understanding of epigenetic modifications and tumour microenvironments, as well as their role and dynamics in CLL evolution.

What do you hope will be the impact of the project on future diagnostics and treatments?

Our hope is to establish cancer evolution as a predictable process. With sufficient understanding of the forces that drive evolution and selective advantages of sub-clonal mutations, we hope to develop prognostic schemes that anticipate individuals' evolutionary trajectories.

Treatments based on these schemes would directly aim to target the cancer plasticity that underlies progression, treatment resistance or relapse. CLL provides us with a unique opportunity to better understand cancer evolution. The conceptual insights about cancer that can thus be gained from CLL would have a high potential for being translated across other haematologic and solid malignancies.

Explore further: Follicular lymphoma: A tale of two cancers

More information: Project page: cordis.europa.eu/project/rcn/186119

Continued here:
CLL evolution under the microscope - Medical Xpress

Yusaku Kodaka1,2, 3, Yoko Asakura1,2, 3, and Atsushi Asakura1,2, 3

1Stem Cell Institute2Paul and Sheila Wellstone Muscular Dystrophy Center3Department of Neurology, University of Minnesota Medical School, Minneapolis, MN

BioTechniques, Vol. 63, No. 2, August 2017, pp. 7276

Supplementary Material

Abstract

Viral vectormediated foreign gene expression in cultured cells has been extensively used in stem cell studies to explore gene function. However, it is difficult to obtain high-quality stem cells and primary cells after viral vector infection. Here, we describe a new protocol for high-efficiency retroviral infection of primary muscle stem cell (satellite cell) cultures. We compared multiple commercially available transfection reagents to determine which was optimal for retroviral infections of primary myoblasts. Centrifugation force was also tested, and a spin infection protocol with centrifugation at 2800 g for 90 min had the highest infection efficiency for primary myoblasts. We confirmed that infected muscle stem cells maintain cell proliferation and the capacity for in vitro and in vivo myogenic differentiation. Our new, efficient retroviral infection protocol for muscle stem cells can be applied to molecular biology experiments as well as translational studies.

Skeletal muscle regeneration is mediated by muscle stem cells called satellite cells (1), which are normally mitotically quiescent in adult muscle. After muscle injury or exercise, quiescent satellite cells undergo activation, followed by proliferation. Proliferating satellite cells, which are myogenic precursor cells, eventually exit the cell cycle and fuse with each other to form multinucleated myotubes. Isolated satellite cells from skeletal muscle can be cultured in vitro as satellite cellderived primary myoblasts (2,3). These primary myoblasts are used for in vitro models of skeletal muscle cell differentiation, self-renewal of satellite cells (4), in vivo satellite cell transplantation (5), and multi-lineage differentiation (6). As opposed to immortalized myoblast cell lines such as C2C12 cells, animal or human primary myoblasts can be utilized for cell transplantation as well as studies of stem cell biology (4,7).

METHOD SUMMARY

Here, we compared multiple commercially available transfection reagents with different infection protocols and determined that a spin infection protocol with centrifugation had the highest infection efficiency for primary myoblasts. The infected cells continued to proliferate and retained the capacity for in vitro and in vivo myogenic differentiation.

One drawback of primary myoblasts is that they need more complex culture conditions to maintain their proliferation and differentiation abilities. The use of high serum conditions for cell growth is an example of this. Furthermore, the efficiency of DNA transfection and viral infection for primary myoblasts is lower than for C2C12 cells (8,9). Retroviral or lentiviral infection has been used for obtaining stable foreign gene expression that enables long-term experiments, including in vivo cell transplantation of myogenic cells (2,10-12). However, the viral supernatant normally contains low levels of nutrients and growth factors, which inevitably induces cell cycle exit followed by myogenic differentiation. Therefore, a method for high-efficiency viral infection without the need for culturing with the viral supernatant is critical for maintaining the ability of primary myoblasts to proliferate and differentiate (13).

For efficient retroviral infection, a spin infection protocol has been established for several cell types, including hematopoietic progenitor cells (14-17). To adapt the spin infection method to primary myoblasts, we identified optimal conditions for both transfection reagents and centrifugation time and force.

All animal experimental protocols were approved by Institutional Animal Care and the Use Committee of the University of Minnesota. Satellite cellderived primary myoblasts such as CD31(-), CD45(-), Sca-1(-), and integrin 7(+) cells were isolated from skeletal muscles of 2 month-old mice (C57BL6, Charles River Laboratories, Wilmington, MA) by MACS separation (Miltenyi Biotec, San Diego, CA) as described previously (3). Myoblasts were maintained on collagen-coated dishes in growth medium (GM) [Hams/F10 (Sigma- Aldrich, St., Louis, MO), 20% FBS, 20 ng/ mL basic FGF (R&D Systems, Minneapolis, MN), and 1% penicillin/streptomycin (Invitrogen, Carlsbad, CA)] (7). Proliferating myoblasts in GM were defined as Day 0. Myogenic differentiation was induced by replacing GM with differentiation medium (DM) [DMEM (Sigma-Aldrich), 5% horse serum, and 1% penicillin/streptomycin] for 3 days.

Retroviral supernatants were produced by transfection of pMX-GFP (Cell Biolabs, San Diego, CA) or a pMX-mCherry retroviral vector into a 293T Platinum-E Retroviral Packaging Cell Line (Plat-E) (Cell Biolabs). One day before transfection, Plat-E cells were cultured in DMEM with 10% FBS and without antibiotics until they reached 70%90% confluency. Various transfection reagents were used: Lipofectamine (Thermo Fisher Scientific, Waltham, MA), Lipofectamine 2000 (Thermo Fisher Scientific), Lipofectamine LTX (Thermo Fisher Scientific), TransIT-293 (Mirus Bio LLC, Madison, WI), TransIT-2020 (Mirus Bio LLC), TransIT-LT1 (Mirus Bio LLC), PolyJet (SignaGen Laboratories, Rockville, MD), and LipoJet (SignaGen Laboratories). Five microliters of each transfection reagent was suspended in 200 l DMEM without FBS and with 5 g of pMX-GFP or pMX-mCherry plasmid DNA for 20 min at room temperature (RT). PlatE cells (6 105) were plated on collagen-coated 3 cm dishes 1 day before transfection. The next day, the medium was replaced with 800 ml DMEM with 10% FBS and 200 ml DMEM using the transfection complex described above. After incubation for 24 h, the medium was changed to 1 mL new DMEM with 10% FBS. Retroviral supernatants were then harvested 24 h after the medium change. Syringe filters (0.45 mm) (Millipore Sigma, Billerica, MA) were used to remove any cells from the retroviral supernatants. Primary myoblasts (1 105) were plated on collagen-coated 3 cm dishes for 24 h before the viral plating infection. Retroviral supernatants were used for viral infection of primary myoblasts with 10 g/mL polybrene (Millipore Sigma) for 4 h, and cells were then cultured in GM for 48 h. For the spin infection, myoblasts were treated with 0.25% trypsin-EDTA (Thermo Fisher Scientific), and 1 105 myoblasts were then transferred into 1.5 mL microcentrifuge tubes. The cells were centrifuged and then resuspended with the retroviral supernatant with 10 g/ mL polybrene. After the myoblast spin infection was performed at RT under appropriate centrifugation conditions, the cell pellets were resuspended with GM and plated on collagen-coated 3 cm dishes. GFP expression was examined 2 days after spin infection. Dead cells were counted by trypan blue (Thermo Fisher Scientific) staining. 5-ethynyl-2-deoxyuridine (EdU) was added to culture plates 3 h before fixation of the cells. EdU staining was performed using the Click-iT EdU Alexa Fluor 488 Imaging Kit (Thermo Fisher Scientific). Immunostaining was performed with anti-GFP (AB3080; Millipore Sigma; RRID:AB_91337), anti-myosin heavy chain (MHC) (MF 20; Developmental Study Hybridoma Bank, Iowa City, IA; RRID:AB_2147781), anti-myogenin antibody (F5D; Developmental Study Hybridoma Bank; RRID:AB_2146602), or anti-phospho-histone H3 antibody (pHisH3) (D2C8; Cell Signaling, Danvers, MA; RRID:AB_10694226), followed by Alexa 488-conjugated anti-rabbit IgG (A-21206; Thermo Fisher Scientific; RRID:AB_2535792) and Alexa 568-conjugated anti-mouse IgG (A10037; Thermo Fisher Scientific; RRID:AB_2534013) or Alexa 488-conjugated anti-mouse IgG (A-21202; Thermo Fisher Scientific; RRID:AB_141607). DAPI (Sigma-Aldrich) was used for counterstaining of nuclei.

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Spin infection enables efficient gene delivery to muscle stem cells - BioTechniques.com