Musculoskeletal Disorders-Stem Cell Based Drug Discovery

A common Neuromics’ theme is harnessing the power of cellsTM. The raw cost of the cells are often the biggest consideration. We encourage our customers to focus on true costs. These include the # of cells (how many times can they be passaged?), culture viability (how long do the cells live) and bioactivity (how closely do cultures mimic in vivo behavior?). I would like to present a presentation and publication confirming our competitive advantage when analyzing true costs.

Setting a higher bar for Neuron-Glial Based Assays!

Dr. Randen Patterson and his team at UC Davis have developed new culturing techniques using our e18 Rat Primary Hippocampal Neurons. They have developed a protocol that allows for culturing of E18 hippocampal neurons at high densities for more than 120 days. These cultured hippocampal neurons are (i) well differentiated with high numbers of synapses, (ii) anchored securely to their substrate, (iii) have high levels of functional connectivity, and (iv) form dense multi-layered cellular networks. We propose that our culture methodology is likely to be effective for multiple neuronal subtypes–particularly those that can be grown in Neurobasal/B27 media. This methodology presents new avenues for long-term functional studies in neurons. This is good news indeed: Todd GK, Boosalis CA, Burzycki AA, Steinman MQ, Hester LD, et al. (2013) Towards Neuronal Organoids: A Method for Long-Term Culturing of High-Density Hippocampal Neurons. PLoS ONE 8(4): e58996. doi:10.1371/journal.pone.0058996.

We will continue to raise the bar. Better cultures=lower costs and better outcomes!

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.

Nanofiber 3-D Cell Based Assays

This “News behind the News” is a historic event.  It demonstrates how nanofiber scaffolds can be used to engineer organs for human transplants. Good news for researchers looking solutions are in vivo like environments for cell based assays.

Nanofibers Solutions work in transplants-imagine how well they will work in your 3-D based cell based assays.

3-D Cell Based Assays for Drug Discovery are the future. Like any new model, adoption rates are a function of how well the new solutions works. “The proof is in the pudding”.

Here’re highlights of a historic event based on transplants using nanofiber engineered laryngotrachea : Collaboration between Nanofiber Solutions and the Karolinska Institutet produces first synthetic laryngotracheal implants seeded with the patient’s stem cells to be successfully transplanted into human patients in Russia.

COLUMBUS, Ohio, June 26, 2012 – Nanofiber Solutions, LLC, an Ohio-based developer, manufacturer and marketer of 3-D synthetic scaffolds to advance basic research, tissue engineering and regenerative medicine announced today the first and second successful transplants of its tissue engineered laryngotracheal implants seeded with cells from the patients’ bone marrow.

The surgeries were performed June 19th and 21st at the Krasnodar Regional Hospital (Russia) by Dr. Paolo Macchiarini, Professor of Regenerative Surgery at the Karolinska Institutet (Stockholm, Sweden), and colleagues. Dr. Macchiarini led an international team that included Dr. Vladimir Porhanov, head of Oncological and Thoracic Surgery at Kuban State Medical University (Russia), Dr. Jed Johnson, Nanofiber Solution’s Chief Technology Officer who created the synthetic organs, Harvard Bioscience (Boston, USA) who produced the bioreactor, and Dr. Alessandra Bianco at University of Rome, Tor Vergata, who performed mechanical testing during scaffold development.

Both patients, a 33 year-old mother from St. Petersburg and a 28 year-old man from Rostov-on-Don, were in au to accidents and suffered from a narrowing of the laryngotracheal junction for which they already had failed previous surgeries. Transplantation was the last option for the patients to have normal quality of life. Immediately following transplantation, both patients were able to speak and breathe normally.

Nanofiber Solutions, lead by Dr. Johnson, designed and built the nanofiber laryngotracheal scaffolds specifically to match the dimensions of each patient’s natural larynx and trachea, while Harvard Bioscience provided a bioreactor used to seed the scaffold with the patients’ own stem cells.  Although this procedure represents the world’s first and second successful use of synthetic synthetic laryngotracheal implants, it is Nanofiber Solution’s second and third successful organ implants using their synthetic scaffolds within the last year.

Nanofiber Solutions’ scaffolds mimic the body’s physical structure and allow for a more successful seeding, growth and differentiation of stem cells. Because the cells used to regenerate the larynx and trachea were the patients’ own, doctors report there has been no rejection of the transplants and the patients are not taking immunosuppressive drugs. (more).

Capabilities of 3-D nanofiber scaffolds for cell based assays:

Human brain tumor biopsy showing migrating tumor cells along the alligned nanofiber.
  • Nanofibers are optically transparent to allow for live-cell imaging and real time quantification of cell mobility using an inverted microscope
  • Nanofibers mimic the 3D topography found in vivo which produces a more realistic cellular response to therapeutics.
  • More realistic cellular behavior means you can use fewer animals and decrease time-to-market for drug discovery and development.
  • Nanofibers can easily be coated with ECM proteins using existing protocols for standard lab ware.
  • Cells can be easily removed for protein or gene analysis using trypsin, EDTA, etc.
  • We will continue posting relevant press releases, pubs and data that prove the capabilities of these important solutions.

    Dr. Jim Musick-Making MSCs Work

    Harnessing the Power of CellsTM

    Dr. Jim Musick

    Dr. Jim Musick

    Dr. Jim Musick and his company, Vitro Biopharma, give Basic and Drug Discovery Researchers the ability to harness the power of Human Mesenchymal Stem Cells (hMSCs). This power is essential for blazing new trails in the stem cell research and regenerative medicine frontier.

    I am pleased to welcome Jim as a partner in providing my company the expertise and knowledge highlighted in this profile. Together, we give our customers, colleagues and friends the ability to easily culture, grow, differentiate and maintain large stocks on hMSCs.

    Background

    Jim received his PhD from Northwestern University in 1975 and then joined the staff of University of Utah where he specialized in the study of Neuroscience and synaptic transmission. He joined UltraPure Laboratories in 1983.

    At UltraPure, he learned the art and science of commercializing biologicals. There he helped develop procedures for the commercial production of purified human pituitary hormones including, prolactin, growth hormone and TSH. This included developing QA/QC procedures to support commercial distribution of these products.

    He joined Vitro Diagnostics in 1988 and directed all operations involved in the establishment of a diagnostic product line that included about 30 different purified antigen products.  His direct responsibilities included research & development, manufacturing, intellectual property development and maintenance, marketing and sales.  He was also responsible for the development & initial commercialization of the fertility drug VITROPIN™ as well as the cell immortalization program of the Company.  He is an inventor or co-inventor of all issued and pending patents owned by the Company.

    In 1998 he also completed an Executive Program at JJ Kellogg Graduate School of Management, Northwestern University in Managing New Product Development.

    In 2000, he orchestrated the sale of the antigen manufacturing division to Aspen Biopharma (Nasdaq, APPY) while retaining IP related to use of FSH as a fertility drug and to cell line generation technology.

    He has the spirit of a polished scientist/entrepreneur with strong operational and process expertise.

    Harnessing the Power of hMSCs

    As President and CEO of Vito Biopharma, Jim leverages his expertise and experience to manufacture Cord Blood Derived hMSCs. The stem cell revolution demands large stocks of cells of the highest quality. Meeting the demand is the key to the development of stem cell related therapies. Vitro Biopharma has the capabilities to delivery.

    It is all about starting with vials of potent and pure hMSCs. From there, the customer can grow and differentiate large stocks and be confident in the quality because Jim’s company has the processes in place to insure this. The cell lines are well-characterized with regard to species authentication using sensitive PCR methods to quantify non-conserved genes including COX1, Cytochrome B and actin.  Vitro Biopharma also utilizes karyotyping to authenticate its  cell lines.  Adventitious agents are also tested negative by sensitive PCR methods including known viral contaminants and mycoplasma.  Performance is assured by rigorous testing of viability, growth rate and differentiation capacity for formation of chondrocytes, adipocytes and osteoblasts.  Finally these cells are characterized with regard to phenotypic cluster designation antigens.

    Current Products

    Native and fluorescent-labeled human MSCs including native and fluorescein/rhodamine-labeled MSC-derived chondrocytes and osteocytes along with MSC-GroTM  growth and differentiation media. MSC-Gro™ media is provided in low-serum, humanized and serum-free formulations for both growth and differentiation.  Humanized serum-free media may be supplemented with allogeneic or autologous serum for direct comparisons of growth and differentiation under these conditions.  Powdered MSC-Gro™ formulations are also provided.  Vitro Biopharma’s human MSCss have the capabilities to be expanded through at least 10 passages at rapid growth rates and can be further expanded to 16 passages (~50 population doublings) at slower growth rates.

    Human MSC-derived Osteoblasts stained with Alizarin red at 100 x.

    Image: Human MSC-derived Osteoblasts stained with Alizarin red at 100 x.

    Vitro Biopharma has recently launched a new and revised website complete with convenient online ordering and detailed product technical information (www.vitrobiopharma.com).

    Futures

    In our interview, Jim gave blinding glimpses of the future especially with regard to new products to extend Vitro Biopharma’s offering of clinical tools to fully explore the ever-expanding therapeutic applications of MSCs. I am excited about the potential. I will keep you posted as new products are commercialized.

    Jim Musick and Vitro Biopharma

    Coming Soon

    Neuromics recently added key products from Dr. Jim Musick and our friends @ Vitro Biopharma. These include potent umbilical cord derived human mesenchymal stem cells and MSCGroTM (best of breed growth and differentiation media). These cells are capable of many passages enabling researchers to build large stocks.

    MSCs_MSCGro

    We will be posting Jim’s profile in several weeks.

    Stem Cell and Cell Based Assays Groups on Linkedin

    I wanted to share some links to groups on Linkedin that have proven a useful resource for me. They are also additive to the stories and data posted here:

    Stem Cell Clinical TrialsStem Cell Clinical Trials

    Note: I am the moderator the the Stem Cell Clinical Trail group and welcome all new members. We are currently 400+ strong and growing.

    3D cell biology : tools & techniques3D cell biology : tools & techniques

    Stem Cell Research Stem Cell Research

    California Institute for Regenerative Medicine (CIRM)California Institute for Regenerative Medicine (CIRM)

    neuropathy and neuropathic painneuropathy and neuropathic pain

    The Gene Silencing Collaboration (RNAi, siRNA, miRNA, Dicer, etc.)The Gene Silencing Collaboration (RNAi, siRNA, miRNA, Dicer, etc.)

    Enjoy.

    Gerry Shaw-Master of World Class Neuronal/Glial Markers

    Build it and They will Come

    Gerry and One of His Triumph's MCs
    Gerry and One of His Triumph’s MCs

    I am pleased to profile Dr. Gerry Shaw, a Professor at the University of Florida and also the Head of EnCor Biotechnology Inc.  His story is a guide for incubating and spinning out a successful biotech company (EnCor Biotechnology, Inc.) from a university research laboratory. It should provide an inspiration for fledgling entrepreneurs as the model required little capital investment and has enjoyed profitable growth.

    The Backstory

    Gerry’s major area of research interest can be summarized as the study of cellular changes resulting from central nervous system damage and disease states. These changes help neuroscience researchers understand the progression and hopefully discover root causes of diseases like Alzheimer’s, Parkinson’s and ALS. Understanding which proteins are involved in particular disease states also has the potential of identifying targets for therapies.

    The story starts with Gerry’s Post Doctoral research at the Max Planck Institute for Biophysical Chemistry in Goettingen, in what was at the time West Germany. Here he joined the world renowned laboratory of Klaus Weber and Mary Osborn. This lab had pioneering several important techniques, notably SDS-PAGE for protein analysis and the use of antibodies in immunocytochemistry. Later, after Gerry left the same lab made key contributions leading to the routine use of RNAi in “knock down” of normal cellular proteins. The lab had developed antibodies to tag the subunit proteins of microtubules, microfilaments, intermediate filaments and other cellular proteins, and then used these antibodies to visualize the proteins in immunofluorescence microscopy and on western blots. This enabled researchers to look at changes in the cellular expression of these proteins in powerful new way. These methods have become vital tools for understanding normal cellular function and what happens when cells transition from healthy to diseased states. This lab was an ideal location for Gerry to learn how to make quality monoclonal and polyclonal antibodies. Good antibody reagents are vital for the correct interpretation of immunofluorescence microscopy and western blots, and he was soon supplying his reagents to friends, collaborators and other researchers all around the world. Success is value as antibodies that do not as work as expected waste research time and resources, while quality reagents soon become appreciated and may get to be standard lab reagents.

    University of Florida

    The University of Florida, in Gainesville imported his expertise when Gerry joined the institute in 1986. Here he continued to make antibodies to Neurofilaments or NFs and other Neuronal-Glial Markers. It’s hard to keep a good thing a secret and Gerry faced growing demand from all over for these reagents. This proved a drain both financially and in terms of time commitment, as well as a significant conflict of interest with his basic biomedical research program.

    MAP2_Doering IHC Image: Co-culture of embryonic mouse hippocampal neurons and astrocytes. Primary embryonic hippocampal neurons at 7 days in vitro, were stained with Microtubule Associated Protein-2 (MAP, green) to enable the visualization of the dendritic arbors. These neurons were cultured on top of a monolayer of primary cortical astrocytes, stained with an antibody directed against

    Glial Fibrillary Acidic Protein (GFAP, red). The cell nuclei were visualized by staining with 4′,6-diamidino-2-phenylindole (DAPI, blue). BMC Image of the Month October 2010

    As a result Gerry took his first entrepreneurial step by selling his most popular reagents in bulk initially to Chemicon (now Millipore-Merck). Like any new business venture, he did not really know what to expect. It should come as no surprise that the reagents sold like hot cakes and the check started rolling in. Other immunoreagent companies approached Gerry and soon he was supplying antibodies to pretty much every major biotechnology vendor.

    ABC Biologicals to EnCor Biotechnology Inc.

    Success breeds success and as sales increased over the 1990s, it was time to form an independent business and so ABC Biologicals Inc. was incorporated in 1999 initially to buy equipment and develop licensing agreements. Since Gerry had income from sales, he was in the unusual and enviable position of not needing grants, investors, loans or cash from any other source, and so could proceed with almost total independence. The company was renamed EnCor Biotechnology Inc. in 2002, and at the same time moved into the Sid Martin Biotechnology Incubator, a lab dedicated to commercialization of intellectual property generated by the faculty of the University of Florida. The University of Florida is unusually experienced at this and is well known for launching Gatorade, Trusopt and many other products. After 4 years EnCor “graduated” from the Incubator and now occupies a facility in Gainesville. The company now has almost 100 products with many more under development. This is good news for the Neuroscience community.

    The EnCor-Neuromics Connection

    Neuromics provides EnCor Biotechnology reagents to researchers studying neuro-degeneration, neuro-regeneration, neuro-development, neural stem cells, mood disorders, brain injury and spinal cord injury. My customers have found EnCor’s reagents to be rock solid and versatile.

    In addition, Gerry and his team have proved adept at culturing our E18 hippocampal neurons and ESC derived hN2TM primary neurons. This is a big plus as we can actually see how the cells and markers could resonate together for use in cell based assays.

    Hippo_MAPT_DC1 Image: E18 hippocampal neurons stained with Tau (red) and Doublecortin (green). The two proteins overlap in the proximal dendrites (yellow) Axons (low doublecortin content) are red. Blue staining is the nuclear DNA.

    Futures

    I am excited by the glimpse of the future that Gerry shared. We can expect many new, novel and important markers in the coming months and years. In addition, he will be manufacturing various Enzyme-linked immunosorbent assays (ELISA). These kits have the potential to help clinicians diagnose the early onset of diseases like ALS, Parkinson’s and Alzheimer’s.

    For example, his company currently sells an ELISA kit for sensitive detection of Phosphorylated Neurofilament-H (pNF-H). Expression of this protein is up regulated in a variety of damage and disease states, and can be used to accurately quantify this up regulation. The kit can also detect pNF-H in the sera and spinal cord fluid (CSF) of animals with spinal cord and brain lesions. This protein is not normally found in sera or CSF, so its presence indicates recent axonal injury as a result of either damage or disease. This suggests pNF-H is a useful biomarker of neuronal and more specifically axonal injury or degeneration, a suggestion supported by a growing list of basic science publications on various animal models and patient types from Gerry’s research lab (e.g. Shaw et al. 2005, Lewis et al. 2008, Boylan et al. 2009, Lewis et al. 2010).

    Given the capabilities of EnCor’s markers, the development of more kits is coming. There could be a day in the not distant future where they give clinicians tools to better diagnose and monitor serious neurodegenerative diseases, leading to better disease treatment and management.

    I will keep you informed on Gerry’s and EnCor’s future developments.

    Coming Soon-Dr. Gerry Shaw

    Zen and the Art of Bio-marker Production

    Up next will be Dr. Gerry Shaw.  Gerry is the founder and head of EnCor Biotechnology, Inc. His company is recognized for creating markers that are engines of Neuroscience and Stem Cell Research.

    Dr. Gerry Shaw with Triumph MC

    Dr. Gerry Shaw with Triumph MC

    I am pleased to represent his company’s reagents. They are well designed, thoroughly tested and proven to work in my customers’ many application.

    They have proven especially effective in working in cell based assays using our eSC derived hNP1 human neurons and e18 primary rat hippocampal neurons.

    Applications include the study of TBI, SCI, ALS, AD, MS and PD.

    Image:  hN2 cells stained with our chicken polyclonal antibody to Vimentin, in red. Islands of Hn2 cells form after 4 days in culture forming beautiful flower like structures. Vimentin is a well established marker of early differentiating neuronal lineage cells. Taken with a 10X objective lens. Blue staining is the nuclear DNA.

    hN2 Cells stained with Vimentin

    hN2 Cells stained with Vimentin

    Harnessing the Power of Neural Stem Cells

    I wanted to share an important presentation by Dr. Steve Stice. He is a featured researcher in “News Behind the Neuroscience News”.

    “Does amplification of neural progenitor cells derived from embryonic stem cells solve problems of cell production and FDA safety standards?”
    Steven L. Stice, PhD
    Professor, GRA Eminent Scholar
    Director of the Regenerative Bioscience Center at University of Georgia
    CSO, Aruna Biomedical Inc.