Network vs Individual Bursting Neurons

Motor Neurons and MEA
Dysregulated bursting is at the root of many motor neuron/neuromuscular junction disease. ArunA Biomedical teaming with Axion Biosystems have generated relevant bursting data from our Mouse Motor Neurons cultured on Axion-Bioystem’s Maestro MEA.

Figure: Mouse Motor Neuron Network Modulation by Bicuculline-ckeck out the entire presentation to learn more: GFP+ Motor Neurons: Development and in-vitro Functional Assessment on Microelectrode Arrays

Protocol User’s Guide for Culturing Motor Neuron on MEA(pdf – 679Kb)

Name Catalog # Type Species Applications Size Price
Motor Neurons-GFP+ Quick Start Kit mMN7205.QS Primary Neurons M Cell Assays 750,000 $349
Motor Neurons-GFP+ HTS Kit mMN7205-HTS Primary Neurons M Cell Assays 4 X 750,000 $989
GDNF (Human, Mouse) PR27022-2 Protein H; M 2 ug 10 ug $108 $205
AB2™ Basal Neural Medium AB27011.3 Cell Growth Media H; M Cell Assays 500 ml $69

We will continue providing you content we believe important. Should you have questions, do not hesitate to contact us. Thank you and we stand ready to serve you and your team.

Pete Shuster-CEO and Owner, Neuromics, 612-801-1007, pshuster@neuromics.com

Neuromics and Vitro Biopharma Partner to offer Human Chondrocytes

More good news from our partnership!

Vitro Biopharma Launches Products for Drug Development & Advancement of Joint Regeneration Technology-GOLDEN, Colo., Oct. 9, 2012 (GLOBE NEWSWIRE) — Vitro Diagnostics, Inc. (OTCQB: VODG), dba Vitro Biopharma, announced the launch of a series of products for use in the development of advanced treatment of injury and diseases of joints. The new products are chondrocytes derived from human adult stem cells, mesenchymal stem cells (MSCs). These cells produce collagen and are critical to proper joint function. Joint disease and injury often involve defects in collagen production and/or damage to chondrocytes due to inflammation, disease or injury. The new products include native and fluorescent labeled MSC-derived human chondrocytes together with SPIO-labeled chondrocytes. The later cells are labeled with super paramagnetic iron oxide (SPIO) and may be used for in-vivo imaging of chondrocytes using MRI (magnetic resonance imaging) a common clinical imaging procedure. The availability of native and multiply-labeled chondrocytes provides customers with numerous options to conduct in-vitro high throughput screening cell assays and to coordinate these studies with in-vivo studies.

Images: Chondrocyte Cultures

Our plans call for the addition of more types of primary cells as well as cells that can be tracked in vivo by MRI. I will keep you posted on these developments. 

Vitro Biopharma and Neuromics Announcement

I recently featured Dr. Musick, CEO of Vitro Biopharma. I am pleased to post our joint announcement on our partnership aimed at Harnessing the Power of CellsTM . Together, we provide researchers potent UCB derived human mesenchymal stem cells and MSGroTM media.

Vitro Biopharma Strengthens and Expands Its Distribution Agreement With Neuromics
Published: July 24, 2012
GOLDEN, Colo., July 24, 2012 (GLOBE NEWSWIRE) — Vitro Diagnostics, Inc. (OTCQB:VODG), dba Vitro Biopharma, announced an expanded relationship with a key distributor of its stem cell-based products, privately-held Neuromics, Inc (www.neuromics.com). Vitro Biopharma officials recently met Neuromics principals at their corporate headquarters in Minneapolis, MN. Neuromics was formed in 2005 by Pete Shuster, a marketing and sales professional with extensive experience in the computer industry who is now focused on providing integral products to the life science industry. Neuromics competitive advantage is based on providing products that lower costs and raise the probability of successful outcomes and include bio-markers, growth factors, transfection reagents, apoptosis detection kits, primary cells and related media. They are designed to serve basic and drug discovery researchers focusing on finding root causes and therapies for neuro-degenerative, autoimmune, inflammatory response diseases and certain forms of cancer.

Vitro Biopharma and Neuromics recently entered a distribution agreement whereby Vitro provides its stem cell-based products to Neuromics’ customers. Stem cell research continues to grow, based on positive results from several studies showing potential clinical benefit of stem cell-based cellular therapy. There is a growing demand for adult stem cells known as mesenchymal stem cells (MSCs). Vitro Biopharma manufactures and sells several fundamental tools needed to advance adult stem cell research and clinical studies, including human MSCs and derivatives, the MSC-Gro™ Brand of media optimized for growth and differentiation of MSCs as well as assay kits for determination of MSC quality, potency and response to toxic agents.

Based on the growing demand for MSC products provided by Vitro Biopharma for both research and clinical studies, Neuromics officials have committed increased resources to promote these products. Furthermore, both Vitro Biopharma and Neuromics are evaluating additional products that are perceived to enhance the competitive advantage of Vitro Biopharma’s existing MSC product line through strategic combination. A strategic goal moving forward, is an additional partner to supply devices needed for FDA-compliant manufacture of clinical materials for use in pre-clinical and clinical studies. These devices are known as bio-reactors that allow for highly controlled and reproducible production of large numbers of stem cells needed to provide therapeutic benefit. Several characteristics of these bioreactors are essential in reaching the demanding goals needed to achieve regulatory compliant manufacturing to support stem cell therapy that maintains rigorous standards of safety and efficacy. We are presently evaluating suitable devices for this application that appear to be well-suited to the required needs, specifications and characteristics.

Dr. Jim Musick, Vitro’s President & CEO, said, “We are very pleased to expand our relationship with Neuromics, Inc and look forward to a long-term mutually beneficial business relationship. While we focus on product development, manufacturing and advancement of our product pipeline, our sales partnerships are important alliances in the expansion of revenue generation. Neuromics is an ideally suited sales partner due to their broad experience and success in distribution of critical life science products. We also see the importance of a complete manufacturing system to support stem cell-based cellular therapy. It is through the highly complex interactions of cells, cell culture media, extracellular matrices and other environmental conditions that together determine the characteristics of the stem cells intended for clinical use. We also anticipate additional expansion of our relationship with Neuromics as Vitro Biopharma’s product pipeline advances through the commercialization process.”

“Neuromics’ strategic direction aligns tightly with Vitro’s goal of ‘Harnessing the Power of Cells™’”, said Shuster, “expanding capabilities enable us to serve an increasing segment of the Biotech and Pharma research community. This is especially true of the stem cell and regenerative medicine research community. Our relationship with Vitro Biopharma adds horsepower to these capabilities. Mesenchymal Stem Cells and Media are raw materials for high throughput and high content cell-based assays. Better materials help insure only the best targets, biologicals and compounds are passed on to downstream processes.”

About Vitro Diagnostics, Inc.

Vitro Diagnostics, Inc. dba Vitro Biopharma (OTCQB:VODG);(http://www.vitrobiopharma.com), owns U.S. patents for production of FSH, immortalization of pituitary cells, and a cell line that produces beta islets for use in treatment of diabetes. Vitro also owns a pending international patent for generation of pluripotent stem cells. Vitro’s mission is “Harnessing the Power of Cells™” for the advancement of regenerative medicine to its full potential. Vitro operates within a modern biotechnology manufacturing, R&D and corporate facility in Golden, Colorado. Vitro manufactures and sells “Tools for Stem Cell and Drug Development™”, including human mesenchymal stem cells and derivatives, MSC-Gro™ optimized media for stem cell self-renewal and lineage-specific differentiation. Vitro recently formed a strategic alliance with HemoGenix®, Inc. (http://www.hemogenix.com/) to jointly manufacture and distribute LUMENESC™ and LumiSTEM™ quantitative assays for determination of stem cell quality, potency and response to toxic agents.

About Neuromics, Inc.

Neuromics (http://www.neuromics.com), located in Minneapolis, MN, is a profitable and growing bio-regents company. The company was initially built by supplying bio-markers to Neuroscience Researchers. Today, Neuromics provides a range of solutions that include markers, growth factors, gene expression analysis tools, apoptosis detection kits, primary cells and related media. These solutions are increasingly being used in combinations by customers to help accelerate or improve the process of drug discovery.

Safe Harbor Statement

Certain statements contained herein and subsequent statements made by and on behalf of the Company, whether oral or written may contain “forward-looking statements” within the meaning of the Private Securities Litigation Reform Act of 1995. Such forward looking statements are identified by words such as “intends,” “anticipates,” “believes,” “expects” and “hopes” and include, without limitation, statements regarding the Company’s plan of business operations, product research and development activities, potential contractual arrangements, receipt of working capital, anticipated revenues and related expenditures. Factors that could cause actual results to differ materially include, among others, acceptability of the Company’s products in the market place, general economic conditions, receipt of additional working capital, the overall state of the biotechnology industry and other factors set forth in the Company’s filings with the Securities and Exchange Commission. Most of these factors are outside the control of the Company.

Investors are cautioned not to put undue reliance on forward-looking statements. Except as otherwise required by applicable securities statutes or regulations, the Company disclaims any intent or obligation to update publicly these forward looking statements, whether as a result of new information, future events or otherwise.

CONTACT:
Dr. James Musick
Chief Executive Officer
Vitro Biopharma
(303) 999-2130 Ext. 3
E-mail: jim@vitrobiopharma.com

Source: Vitro Diagnostics, Inc.

Dr. Ivan Rich and HemoGenix

Stem Cells Testing Tools that enlighten Drug Discovery and Cell Therapy Researchers
I am pleased to profile Dr. Ivan Rich. He is the founder, chairman and CEO of HemoGenix and an internationally recognized leader in hematology.  I am timing this profile to coincide with Neuromics launch of HemoGenix’s first to market fully standardized, proven and cost effective  ATP-based, in vitro bioluminescence and high-throughput screening (HTS) cell based assay systems.

These assays represent best in class solutions for detecting and measuring cell viability, functionality, growth, proliferation and cytotoxicity of stem and progenitor cells for stem cell and basic research, cellular therapy, in vitro toxicity testing and veterinary applications.

Hemogenix_Pic

 ivan-rich

2000-Present- Hemogenix-CEO
and Chairman

1996-2000-Palmetto Richland Memorial Hospital

 1995-Second Thesis in Experimental Hematology, University of Ulm

1981-1983-Post Doc University of Chicago

1973-1978-Ph.D. University of Ulm, Biology

 

Ivan’s journey leading to founding of HemoGenix provided him a unique blend of scientific, entrepreneurial and operational expertise.  These traits are the drivers that enable him to invent, successfully commercialize and continuously improve cell based assay systems. These systems meet a wide range of demanding requirements. These include, for example, meeting the requirement by Standards Organizations and Regulatory Agencies for “appropriate” and “validated” assays that can be used by cord blood banks and stem cell transplantation centers to determine whether a stem cell product has the necessary potency characteristics and can be released for transplantation into a patient…high standards indeed!

The Back Story-Hematology and Hemopoietic Stem Cells

Ivan received his PhD from the University of Ulm, in Germany in 1973 in Human Biology. He then completed a second thesis in 1995 in experimental hematology.  Our story starts here.  As a background we need to understand:  the hemopoietic stem cell compartment consists of cells which are responsible for maintaining the steady-state production of some two million red blood cells and two hundred thousand white blood cells every second of a person’s life!

Beginning in 1973, he worked extensively with “classic” colony-forming cell (CFC) assay.  At the same time, He also gained experience in culturing erythropoietic progenitor cells (BFU-E and CFU-E) under low oxygen tension. His group was the first to demonstrate that macrophages grown in vitro could respond to low oxygen tension by regulating erythropoietin production at a local level. His group also demonstrated the role of HOXB6 in erythropoietic development as well as the role of the Na/H exchanger in hematopoiesis. “Necessity being the mother of invention”, Ivan began developing these assays into miniaturized format.  Assays necessary for fully understanding the potential and associated risks of using of these cells for human therapies.

This opened the door for him to do a post doc with the late Dr. Eugene Goldwasser at the University of Chicago. Dr. Goldwasser was renowned for discovering the first partial amino acid sequence of erythropoietin (EPO). This discovery eventually led to the production of human recombinant EPO by Amgen and the development of first EPO related therapeutic (Epogen). It is used to treat anemia from kidney disease and certain cancers.

We now move to Palmetto Richland Memorial Hospital in South Carolina where Ivan served as Director of Basic Research for Transplantation Medicine. From this research,  we learn that the most primitive stem cells have the greatest potential for proliferation and long-term reconstitution of the hemopoietic system, while the most mature stem cells have only short-term reconstitution potential. These primitive cells then become the most excellent candidates for future therapies. BUT how do we know the population of cells derived from cord blood or bone marrow contain the required population of potent and safe (phenotypically stable) primitive stem cells for effective therapies? We can ask the same questions for other stem cell populations that are candidates for therapies. These include mesenchymal stem cells, neural stem cells and others.

Introducing Quantitative, Accurate and Proven High Throughput (HTS) Stem Cell Assays

Ivan and HemoGenix began answering these questions in 2002 with help from National Cancer Insitute (NCI) SBIR grants. This led to the successful launch of the HALO® family of kits. These kits are based on Bioluminomics™ which is the science of using the cell’s energy source in the form of ATP (adenosine triphosphate) to provide us with a wealth of information. The production of ATP is an indicator of the cell’s cellular and mitochondrial integrity, which, in turn, is an indicator of its viability and cellular functionality. ATP also changes in proportion to cell number, proliferation status and potential, its cytotoxicity and even its apoptotic status.

HemoGenix continues to develop and evolve kits key to developing effective and safe stem cell related drugs and cell based therapies.

Practical Applications

Here are examples of the kits in action.

  • HemoGenix and Vitro Diagnostic-Via this partnership, LUMENESC kits for mesenchymal stem cells include high performance growth media for research, quality control or potency or cytotoxicity to the mesenchymal stem cell system
  • LumiSTEM™ for testing  hNP1™ Human Neural Progenitors Expansion Kit-enables  fast, accurate and multiplex detection system for hastening advances in drug safety and discovery as well as environmental toxicology. . LumiSTEM™[now LumiCYTE-HT]  kits are used for in vitro detection of liver toxicity, with an overall reduction in drug development cost for drug candidates
  • High Throughput (HTS) Screening of Multiple Compounds using HALO®-(to learn more see: TOXICOLOGICAL SCIENCES 87(2), 427–441 (2005) doi:10.1093/toxsci/kfi25). Eleven reference compounds from the Registry of Cytotoxicity (RC) and eight other compounds, including anticancer drugs, were studied over an 8- to 9-log dose range for their effects on seven cell populations from both human and mouse bone marrow simultaneously. The cell populations studied included a primitive (HPP-SP) and mature (CFC-GEMM) stem cell, three hematopoietic (BFU-E, GM-CFC, Mk-CFC) and two lymphopoietic (T-CFC, B-CFC) populations. The results reveal a five-point prediction paradigm for lympho-hematotoxicity.
HSC Toxicity Data

HSC Toxicity Data

Futures

The dawn is breaking for stem cells therapies. These cells are the reparative engines for damaged cells in our bodies. These therapies have the potential to alleviate the world’s most insidious, chronic and costly diseases. Tools that enable us to understand the true properties and potency of these cells lower the cost of discovering drugs and cell based therapies.

I look for more tools to spring from the vision of Dr. Ivan Rich that will play an ever increasing and important role in the world of basic stem cell research, stem cell based therapies and regenerative medicine. I plan to keep you updated on the evolution and capabilities of these inventions.

Satish Medicetty-Platforms for MS Drug Discovery

In Search of Remyelination Therapies

Multiple Sclerosis (MS) is an inflammatory disease with no known cure. It affects over 400,000 people in the US and over 2.5 million people worldwide and is the leading cause of non-traumatic neurological disability in North America.

It is a chronic and brutal disease that attacks the brain and spinal cord. MS symptoms are due to the damage or loss of myelin sheath that surrounds, isolates and protects axons of brain and spinal cord. The results are often debilitating and afflict most sufferers in the prime of their lives. The annual costs to slow the disease and treat related
symptoms are in the billions of dollars and rising. There are currently no therapies to reverse damage of MS. At this point, there are only immune suppressive therapies that slow attack on the myelin sheath.

It is with hope and optimism that I present Dr. Satish Medicetty and his company, Renovo Neural Inc. (RNI) in this edition of the “News Behind the Neuroscience News”.

I became aware of Satish and his company in my search for Stem Cells that would broaden Neuromics ability to serve early phase Neuroscience Drug Discovery.

Satish Medicetty

Satish Medicetty

Apr 2010 – Present: President and Board Director Renovo Neural Inc.

June 2008 – Mar 2010: Director of Stem Cell Research and Lab Operations
NeoStem Inc

July 2005 – June 2008: Senior Scientist Athersys

2006 – 2008: MBA, Case Western University

2002 – 2005: PhD, Kansas State University

After my first conversation with him, I was impressed with the capabilities RNI offered.

RNI

The company, founded in 2008 with a$3 million grant from the State of Ohio’s Third Frontier Commission, is leveraging cutting edge research from Dr. Bruce Trapp’s lab at the Cleveland Clinic.

At the core, RNI offers pioneering and propriety assays that give Drug Discovery Companies the ability to screen small molecules and compounds that could be lead therapy candidates for MS and other myelin-related diseases. These screens use a type of stem cell called adult oligodendrocyte precursor cells (OPCs).

The Power of OPCs

So what makes these OPCs an engine for finding cures for MS?  Inflammation associated with MS attacks destroys cells called oligodendrocytes that produce myelin. The only way to reverse this autoimmune related process is for the brain to produce healthy cells that can catalyze re-myelination. Enter OPCs.

OPCs are the raw material for processes the central nervous system uses to manufacture oligodendrocytes.  The brain’s inability to produce enough healthy cells to keep up with the destruction is a root cause of the disease. Understanding how to kick start and keep the oligodendrocyte factory running is a key to reversing this relentless destruction.

Delivering Value

RNI has the capabilities to the decrease time required and increase the odds for discovering potential MS therapies.  They have the raw material (OPCs) and the know how to encourage their transformation into myelinating cells. This expertise can be utilized can be used then to rapidly test new compounds both in vitro and in vivo.

In Vito Assays Example

In Vitro Assays Example

The features of their in vitro assays include:

  • Stringent protocols to generate relatively homogeneous (>85% pure) and consistent population of OPCs as a reliable starting material for HCS assays
  • Relatively high throughput primary screen to identify potential candidates that promote OPC proliferation and/or differentiation
  • Secondary screen to confirm and qualify compounds for further pharmacological testing
  • Positive and negative controls that demonstrate the utility of HCS assays to identify lead candidates that promote OPC proliferation and differentiation.

The features of their in vivo cuprizone assays include:

  • Stringent protocols to generate highly reproducible demyelination/remyelination cuprizone model
  • Cuprizone model recapitulates the in vivo process of demyelination and remyelination in the brain.
  • Cuprizone model provides consistent and accurate results for key regions of the brain that are affected in MS patients including both white and gray matter regions – corpus callosum, hippocampus and cortex.
  • Proof of concept studies demonstrate the utility of our in vivo remyelination assays to evaluate preclinical efficacy of potential remyelination therapies

The end goal is to discover therapies that repair neurons damaged by MS via remyelination and to get them in the hands of people that need them. I will keep you posted on their progress.

25 Best Blogs for Following Stem Cell Research

Providing research proven and reasonably priced Stem Cell Research Reagents is core to our business growth.  Part of my business strategy includes keeping the Stem Cell research community up to date on latest news, methods and publications. This helps oil the engines of basic research and drug discovery.

hN2 Cell-Differentiation

Images Courtesy of Paula M. Keeney, Laboratory and Research Manager, VCU Parkinson's Disease Center of Excellence.

This listing comes to me from my friend Roxanne McAnn at Nursingdegree.net.

Stem cell research has been a contentious issue in both the scientific and political spheres for quite some years. Despite the ongoing battle between those who support and those who oppose the research and treatments, new discoveries and advances in the field are being made all the time. Whether you’re pursuing a career in medicine or science, if you’d like to keep up with these advances, then blogs on the issue are one of the best tools out there. Here, you’ll find a collection of blogs that provide all the information you’ll need to stay on top of the latest in stem cell discoveries.

News-These blogs will let you stay on the cutting edge of new developments in the stem cell research community.

  1. The Stem Cell Blog: Through this blog, you’ll be able to get updates on the latest and greatest in stem cell research.
  2. Stem Cell News Blog: This blog collects a wide range of articles related to stem cell treatments, research and policy.
  3. Ben’s Stem Cell News: Ben Kaplan is a stem cell activist, blogger and a biotech professional who shares his thoughts and the latest information on stem cells here.
  4. Stem Cell Directory: No matter what kind of stem cell information you’re looking for, you’ll find it here through articles, news and videos.
  5. All Things Stem Cell: From treating baldness to cancer, learn about the myriad of ways stem cells may be able to help patients on this blog.
  6. Cell News: This blog will make it simple to be in-the-know when it comes to everything related to stem cells.
  7. The Stem Cell Trekker: Use this blog to learn more about stem cell innovations around the globe.
  8. StemSave: You might not think dental care when you think of stem cells, but this blog will show you that stem cells may be able to be taken from the teeth, giving you a whole new appreciation for those chompers.
  9. Joescamp’s Stem Cell Blog: This blog offers up news, information and insights into adult stem cell research.

Businesses and Organizations-Check out these blogs to see what research corporations and organizations
invested in stem cells are doing.

  1. International Stem Cell Corporation: Visit this blog to learn more about stem cell research that’s being done overseas, as many countries don’t have the same restrictions on research as the U.S.
  2. ViaCord Blog: This company, invested in cord blood baking and research, shares advances in the field of stem cells and cord blood treatments through this blog.
  3. Stem Cell Network Blog: Based out of Canada, this organization’s blog will help readers stay on top of new studies being done in the field, as well as some political issues that will affect researchers in Canada and around the world.
  4. Stem Cell Aware: Here you’ll find articles and information that can help you learn more about individuals who are receiving treatment with adult stem cells around the world.
  5. Umbilical Cord Blood Blog: Learn more about donating umbilical blood and the stem cell research being done with it through this organization’s blog.

Commentary Here, you’ll get not only news, but commentary on stem cell issues as well.

  1. David Granovsky’s Stem Cell Blog: Ranked as one of the top health bloggers by Wellsphere, David Granovsky’s blog on stem cells is sure to provide you more  information on the subject than you’ll have time to read.
  2. California Stem Cell Report: See how stem cell politics are affecting research and development in California through this blog written by journalist David Jensen.
  3. Advance Stem Cell Research: Follow the latest news and commentary on stem cells with this blog.

Research-These blogs, many from labs and experts in the field, focus on providing news and information on the best research being done with stem cells in the world.

  1. Knoepfler Lab Stem Cell Blog: The UC Davis School of Medicine maintains this blog, providing readers with information on everything stem cell as well as other science-related issues.
  2. CIRM Research Results: The California Institute for Regenerative Medicine shares their latest discoveries and political battles here.
  3. Robert Lanza, MD: Dr. Robert Lanza is a scientist and professor working on issues related to cell technology and engineering; his blog will provide readers with some insights into the field and his research.
  4. Stem Cell Gateway: Whether you live in the U.S. or abroad, this blog is the place to visit for information geared towards the stem cell research community.
  5. Tissue and Cellular Innovation Center Blog: Focused on tissue engineering and stem cell biology, this center is at the forefront of much of the research they share via this blog.
  6. Stem Cell Breaking Research: Need to know the absolute latest on stem cell research? This blog may be one of your best bets, with updates posted every day.
  7. Stem Cell Digest.net: On this blog, you’ll find information about stem cell research, progress, new applications and companies who are doing the work.
  8. Stem Cell Methods: Researchers, scientists and medical professionals can learn more about the protocols and methods being used in stem cell research and treatment through this blog.

Author’s not (6/1/2011). This excellent site was brought to my attention by Dr. Anthony G. Payne- www.stemcelltherapies.org: This site is run by Steenblock Research Institute (San Clemente, California) which is a 501(c)(3) non-profit organization devoted to stem cell related education and research (SRI has a massive library facility and  stem cell R & D laboratory).

Ion Channels and Neuromics’ STEMEZ Cells

In my conversation with neuro-drug discover researchers, I am frequently being asked about the potential of using our STEMEZ(TM) hNP1 Human Neural Progenitors Expansion Kits for studying ion channels. How effective are these cells as a source for studying neurodegenerative diseases and for drug screening assays?  There is good news from Dr. Steve Stice and my friends from ArunA and UGA.

When differentiated, these  neural progenitors express subunits of glutamatergic,  GABAergic, nicotinic, purinergic and transient receptor potential receptors. In addition, sodium  and calcium channel subunits were also expressed. Functionally, virtually all the NP cells exhibited delayed rectifier potassium channel currents and some differentiated cells exhibited  tetrodotoxin sensitive, voltage-dependent sodium channel current under whole-cell voltage clamp and action potentials could be elicited by current injection under whole-cell current clamp.  These results indicate that removing basic fibroblast growth factor from the neural progenitor cell cultures leads to a post-mitotic state, and also results in the capability to produce excitable cells that can generate action potentials. This is the first data demonstrating capabilitiesof these cells for ionotrophic receptor assays and ultimately for electrically active human neural cell assays for drug discovery.
hNP1_Gene_Expression

Images: Glutamate receptor expression in hNP cells and differentiated hNP cells The expression of ionotropic glutamate receptors might also be an indicator of neuronal maturation. These receptors are composed of three distinct families: NMDA, kainate and AMPA receptors. The hNP cells and differentiated hNP cells cultured in the absence of bFGF for 2 weeks were analyzed for mRNA expression of subunits of each glutamate receptor subtype relative to hESCs. Significant increases (p<0.05) in Grin2b were seen in hNP cells (20 fold) and differentiated hNP cells (25 fold) relative to hESCs (Figure 3A). Additionally, Grin1 and Grin2d were significantly increased (p<0.05) only in differentiated hNP cells relative to hESCs, but not in undifferentiated hNP cells (Figure 3A). Of the kainate receptors, Grik4 and Grik5 were significantly (p<0.05) increased only in undifferentiated hNP cells relative to hESCs (Figure 3B); whereas, Grik2 was significantly (p<0.05) increased only in hNP cells where bFGF had been removed (Figure 3B). AMPA receptor subunits were also examined. Gria1 and Gria4 were up regulated in hNP cells relative to hESCs (Figure 3C). Two week differentiated hNP cells showed significant (p<0.05) up regulation of Gria2 and Gira4 relative to hESCs (Figure 3C). To determine if functional glutamate channels exist in differentiated hNP cells, calcium influx in response to AMPA, kainic acid or NMDA application was measured on hNP cells, 14 days after the removal of bFGF. Figure 3G indicates that NMDA could not depolarize differentiated or undifferentiated hNP cells enough to cause significant calcium influx above background. In contrast, AMPA and kainic acid can cause calcium influx which can be potentiated by AMPA receptor specific modulator, cyclothiazide (50 μM, Figure 3G).Calcium influx was detected in the presence of cyclothiazide in calcium activity as measured (Figure 3H).

hNP1_Electrophysiology

Images: Sodium channel activity in differentiated hNP cells was measured using whole cell voltage clamp. 81 total hNP cells cultured in the absence of bFGF from 4 to 27 days were analyzed. Of these, 34 exhibited no fast inward currents in response to a step depolarization indicating the 348 absence of functional voltage gated sodium channels (Figure 4G). The remaining cells yielded between 0.04 – 1.5 nA of inward current in response to the step depolarization (Figures 4B and 4G). These currents inactivated rapidly in all cases (Figures 4B and 4C) and could be abolished with the addition of 1 μM TTX (n = 3 cells; Figure 4C). Voltage-dependent steady state inactivation (n = 11 cells; Figure 4D) and recovery from fast inactivation (n = 5 cells; Figure 4E) were also observed on several positive cells. A subset of these cells was subjected to current clamp and action potentials were elicited by current injection (n = 8 cells, Figure 4F). In support of this, increasing concentrations of a sodium channel activator veratridine in a FLIPR assay on differentiated hNP cells show an increasing calcium response (Figure 4H). This probably resulted from voltage-gated sodium channel depolarization of cells that subsequently allowed calcium influx through calcium channels. These data indicate that differentiation of hNP cells by removal of bFGF can lead to a neuronal cell that can generate action potentials and depolarize the cell. The 58% hit rate for voltage-gated sodium channel function (Figure 4G), does not reflect the true proportion of sodium channel positive cells in our differentiated hNP cells, but rather our ability to morphologically distinguish these cells from negative cells by eye. An example of the morphology of a sodium channel positive cell is shown in Figure 4A. The positive cells were phase bright with a few long processes.

STEMEZ hNeural Progenitors and Cell Migration

I first featured Dr. Steve Stice in August 2008. I have since done follow up posts based on the excellent studies they have been conducting using our  STEMEZ (TM) Human Neural Progenitor & Neuron Discovery Kits.

I would like to highlight a poster based on research Steve and his Team conducted with Platypus Technologies.

Allan C. Powe, Jr., Kathryn L. Hodges, Jamie M. Chilton, Scott Gehler, Renee L. Herber, Keren I. Hulkower, Steven L. Stice. Identification of stimulators and inhibitors of cell migration in human embryonic stem cell derived neural progenitors using a novel, high throughput amenable assay platform.

Investigates the migratory behavior of an adherent monolayer neural progenitor cell line derived from human embryonic stem cells (hNP1 ™; ArunA Biomedical)using a novel 96‐well based cell migration assay platform (Oris™ Cell Migration Assay; Platypus Technologies) amenable for high throughput screening. The assay platform uses stoppers to create central exclusion zones within the wells; cells are plated outside the zone and migrate inward once the stopper is removed.

Data suggest this is a tool for understanding proper nervous system development, development of therapies for cell migration defects, and identifying novel environmental neurotoxicants.

Conclusions:
—The hNP1™ Oris™ Cell Migration Assay can quantitatively detect both stimulators and inhibitors of cell migration.
—Method development to date indicates that the assay has the potential for adaptation as a homogenous HTS‐suitable cell‐based assay.
—Preliminary results suggest that bFGF alone has a potent chemokinetic effect while LIF and GDNF act synergistically to drive migratory behavior during dopaminergic differentiation.

Dr. Steve Hall knows Stem Cells

 
Developing New Methods, Applications and Reagents for Regenerative Medicine and Stem Cell Research.
 Getting Started with Alphagenix                    

Steve is an advisor, collaborator and friend. He has the innate ability to bring his his scientific expertise and entrepreneural insticts together in a way that anticipates emerging needs of the research community we both serve. He is an expert in immunology, neuroscience, virology and regenerative medicine (stem cells).  Most notably, he is the sole inventor on the patent that formed the basis for using the Nodaviruses as vaccine and gene therapy vectors U.S. Patent 6,171,591. These vaccines are in various stages of preclinical development as are protoype therapeutic vaccines for neurodegenerative diseases. 

Our two companies first worked together to identify and manufacture several important stem cell markers. We tested potency on our STEMEZ hNP1TM Human Neural Progenitors. They proved to be effective. This confirmed Steve’s ability to identify, design and make these markers. The demand for them continues to grow.

These successes were a prelude of good things to come.

Current Focus

Steve is currently  developing novel products and technologies for basic and clinical research with a particular emphasis on stem cell markers, biomaterials and regenerative medicine. The biomaterials product focus involves the design and application of 3-dimensional biomaterials comprised of extracellular matrix components and peptide nanofibers that have cell culture and tissue engineering applications. In addition, the company conducts regenerative medicine research that involves basic science and translational preclinical research using stem cell regulatory network discoveries and novel preclinical studies utilizing animal models with a focus on neurological disease and diabetes.

He is a contributor to: Stem Cell Therapy for Neurological Diseases Stem cell therapy for the treatment of acute and chronic neurological diseases

Dr. Steve Hall

2001-Present-President-Alphagenix

2006-2007-CSO-Neuromics

2000-2001-President-AmProx, Inc

1996-2004-President and CSO-Pentamer Pharmaceuticals

1996-1997-Sr. Research Fellow-Medical Biology Institute.

1995-1997-Research Associate-Scripps Research Institute

1995 PhD Purdue University

Musashi-1 Antibody

Musashi-1 Antibody

Image: Musashi (green) staining of neural rosettes(human). Nuclei are counterstained blue (DAPI). Image courtesy of Dr. Steve Stice and Dr. Patricia Wilson, University of Georgia.

 

 

Harting, Matthew T., Cox, Charles S. and Hall, Stephen G.  Adult Stem Cell Therapy for Neurological Disease: Preclinical evidence for cellular therapy as a treatment for neurological disease. In Vemore and Vinoglo (eds): Regulatory Networks in Stem Cells. Humana Press, pp 561-573, (2009). More information.

Specific Projects

  1. Steve has 3 major projects underway:
    In collaboration with Dr.  Charles Cox , Distinguished Professor, UT Medical School @ Houston, Steve has been using stem cells to treat  Traumtaic Brain Injury (TBI) in Rat. Neural stem cells transplanted into the site of injury. In this model, treated rats showed injury significantly improved motor skills with a moderate recovery in cognitive ability. This research forms the base for eventually repairing damage in humans suffering TBI. Methods and reagents developed also could be useful for basic research and drug discovery.
  2. Steve is working with Burnham Institute to develop methods for using  Human Mesenchymal Stem Cells to regenerate beta cells. This research holds promise for type 1 diabetics.
  3. Steve developing biomaterials including extracellular matrix proteins in novel cell culture systems and synthetic peptide nanofibers for these purposes.  It is investigating stem cells and genetically engineered cells and their interaction with these biomaterials, which has the ability to increase the efficacy of cell therapy. This is highlighted by a human laminin sytem that shows promise in restoring function in Muscular Dystophry.

The last project is promising enough that it could lead to funding for phase 1 testing.

I will continue to keep you posted on progress. I am excited about the new regeants and method that evolve from Steve’s Research. As these prove to work in unique and novel ways, the will become available to Neuromics.

Coming Soon-Dr. Steve Hall

Dr. Steve Hall

Dr. Steve Hall

Dr Steve Hall has been a friend, collaborator and mentor since I purchased Neuromics. This includes being a Neuromics’ Premier supplier of Stem Cells and Related Markers, Media and Methods. Steve is currently President at AlphaGenix, Inc.

His expertise includes developing novel products and technologies for basic and clinical research with a particular emphasis on stem cell markers, biomaterials and regenerative medicine. The biomaterials product focus involves the design and application of 3-dimensional biomaterials comprised of extracellular matrix components and peptide nanofibers that have cell culture and tissue engineering applications. In addition, the company conducts regenerative medicine research that involves basic science and translational preclinical research using stem cell regulatory network discoveries and novel preclinical studies utilizing animal models with a focus on neurological disease.

He is a contributor to: Stem Cell Therapy for Neurological Diseases Stem cell therapy for the treatment of acute and chronic neurological diseases

Harting, Matthew T., Cox, Charles S. and Hall, Stephen G.  Adult Stem Cell Therapy for Neurological Disease: Preclinical evidence for cellular therapy as a treatment for neurological disease. In Vemore and Vinoglo (eds): Regulatory Networks in Stem Cells. Humana Press, pp 561-573, (2009). More information.

Stay tuned for Steve’s backstory in June!