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. 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.

    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).

    Studying Apoptosis In Tumors

    I have featured Gary Johnson here.

    I value my partnership with his compant, ICT. They provide our customers with potent and research proven Apoptosis Kits and Methods. Here we feature publications referencing our MitoPT™ Kits. These Kits easily assess changes in mitochondrial membrane potential. Changes in mitochondrial membrane potential can correlate with cytochrome c release and the initiation of apoptosis.
    A431 cells, treated with predetermined IC50
    concentration of novel anticancer agents, fluoresce green and orange-red with MitoPT JC-1. Data courtesy of Zayas/ Carro, Universidad Metropolitana.
    Anticancer Effects of Alpinia pricei Hayata Roots.
    CL Hsu, YS Yu, GC Yen. J. Agric. Food Chem., Jan 2010, 58 (4), pp 2201–2208.

    Anticancer Effects of Flavonoid Derivatives Isolated from Millettia
    reticulata Benth in SK-Hep-1 Human Hepatocellular Carcinoma Cells.

    SC Fang, CL Hsu, HT Lin, GC Yen. J. Agric. Food Chem., Jan 2010, 58
    (2), pp 814–820.

    Mechanisms of Apoptotic Effects Induced by Resveratrol,
    Dibenzoylmethane, and Their Analogues on Human Lung Carcinoma Cells.

    CJ Weng, YT Yang, CT Ho, GC Yen. J. Agric. Food Chem., Jun 2009; 57
    (12), pp 5235–5243.

    Gary Johnson-Apoptosis Ace

     Gary Johnson

    1994-present-President, ICT

    1993-1996-Conjugation Chemist, R&D Systems

    19989-1993-Supervisor Protein Conjugation & ELISA Development Group, Solvay Animal Health

    1986-1989-Immunologists, Biosciences Lab, 3M

    1976-1986-Various Lab, U of MN

    Gary’s Conatct Info:

    Inventing Better Ways to Measure Apoptosis 

    This profile features another Scientist Entrepreneur. Dr Gary Johnson is the Founder and President of Immunochemistry Technologies LLC (ICT). His company manufactures kits that have the capabilities to quantitatively measure apoptosis effects. This is important to Neuromics, because these are core to many diseases of research interest to our customers. These range from Cancer where apoptosis detection can be used to to visualize the efficacy of tumor killing therapies to Neuroscience where apoptosis could be a root cause of many cognitive and neuro-muscular diseases.

    I am excited about featuring Gary. I have been working with him and his team over the past 5 years. They have actively supported my company in providing Apoptosis Research Kits. The strength in our relationship is built on his company supplying best of breed reagents. The feedback I receive from users is overwhelmingly positive. In addition to these kits, ICT is also recoginized for their rock solid ELISA Buffers and Diluents.

    It takes a unique blend of business and scientific acumen to build a company like ICT. So let’s start with Gary’s background and experience and then on to the specifics on his company and products and what sets ICT apart from competitors.

    Gary’s Background

    Gary’s began his career at the University of Minnesota in 1978 where he worked in a variety of labs. There he gained a wealth of experience and expertise in research techniqes. These included chromatography, immunoelectrophoresis, radiolabeling, mass spectrometry,  proton NMR spectroscopy and western blotting.

    He leveraged his abilities and became more deeply involved in immunobiology. He  joined Dr. Harry Orr’s lab in 1981. There he used recombinant DNA techniques to study the class I genes of the major histocompatibility complex and he also supervised the tissue culture work. This provided the stepping stone to Dr. David Klein’s lab in 1984. There he studied the difference between diabetic and non-diabetic glomerular basement membrane proteoglycans in kidney disease. In order to do this research Gary developed in vivo or in vivo labeling techniques.

    Gary then moved from University to commercial labs. We will see how his growing expertise morphed into the founding of ICT and why his broad knowledge and experise enabled a successful launch of the company.

    From 1986 until founding ICT Gary worked at 3M, Solvay Animal Health and R&D Systems. Over his tenure, he worked as an Immunologist, Supervised an ELISA and Protein Purification and was a Conjugation Chemist. Having mastered a unique range of basic and commercial bio-research techniques, the evolution to Scientist-Entreprenuer was a natural next step.

    In 1994, Dr. Brian Lee and Gary launched ICT. The company’s early success was in contract assay development. The revenue generated from these programs, has enabled ICT to manufacture and release a growing catalog of Apoptosis Detection Kits.

    ICT’s Products and Capabilties

    ICT’s provides proprietary probes for measuring apoptosis in vitro and in vivo. These probes are used by researchers  to detect caspases, cathepsins, serine proteases, cholinesterase enzymes, and assess mitochondrial health.Applications include: assessing the efficacy of chemotherapy, to quantifying  neurodegeneration, and early detectionof eye disease, to name a few.

    Specific Products Include:

    keratconus1

    Images: Normal (left) and keratoconus (right) corneal fibroblasts were labeled with Caspase 3 & 7 Assay Kit, green.

    Pacing the Field

    ICT is setting the pace in Apoptosis Detection by  recognizing and resolving issues inherent in competitive offerings. These include:

    1. Difficulty permeating cells.
    2. High background problems.
    3. Does not bind to early stage apoptotic cells.
    4. Not as sensitive as a cell permeant inhibitor probe.
    5. Does not bind to all apoptotic tumor cells (Dicker, Cancer Biol. Ther., 2005. 9:1014-1017).
    6. Binds positively to normal and healthy bone marrow derived cells (Dillon, J. of Immunol., 2001. 166:58-71).
    7. Many in vitro protocols involve lysing the red blood cells before running flow cytometry, this method results in the binding of Annexin V to all of the cells in the sample (Tait, Blood, Cells, Molecules, and Diseases., 1999. 25:271-278).  The inversion of PS and cells containing large amounts of PS may not be related to apoptosis and this adds to the background issues.
    8. Does not measure a process of apoptosis, but rather an effect of apoptosis.

    Capabilities that will enable them strengthen their leadership position include:

    1.  Uses a cell permeant probe that can easily penetrate tissues and cells.
    2. Very sensitive.
    3. Specific, no reported false positives.
    4. It is a direct measurement of an intracellular process of apoptosis, detects only active caspases and caspase active cells are always apoptotic.
    5. Passage through the blood-brain barrier has been demonstrated.
    6. Passage through the blood-retinal barrier has been demonstrated.
    7. No background problems when injected intravenously.
    8. Detects very early through late stage apoptosis.

    ICT is continuing to invest heavily in developing new capabilties. Gary highlighlighted some of the breakthroughs that are on the horizon. I plan on announcing these as they become public.Stay tuned.