Wednesday, June 27, 2012

Laryngotracheal Transplantations Using Nanofiber Solutions Scaffolds

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.

    Sunday, June 24, 2012

    100% Animal Free FGF Basic

    New ISO-kineTM FGF basic-only 39 USD-10 ug.

    Fibroblast Growth Factor (FGF) Recombinant Proteins are widely used as tools in Stem Cell Research. They have the capabilities to catalyze growth and differentation as well as maintain cell stasis. We continue to offer new FGFs to make sure we meet the unique demands of our cutsomers and collaborators  .

    Our E. Coli derived Human FGF (146 aa) recombinant protein is our #1 seller. As Stem Cell research moves from the bench top to the bedside, there will be a growing need for 100% animal free proteins. We are please to announce we have added a 100% animal free FGF (146 aa) basic that has bio-activity comparable to our top selling options
    Image: ISOKineTM bFGF vs our potent and proven e-coli derived bFGF.

    I anticipate offering more ISOKineTM Bio-risk free Stem Cell Research growth factors to meet growing demand. This growth will be driven by their unique advantages. These include low protease activity and secondary metabolite content and simple protein content all of which aid in downstream processing. These products also have the G.R.A.S. (Generally Recognised As Safe) status from FDA.

    Wednesday, June 20, 2012

    TRPA1 and Tooth Pain

    Our TRPV (Vanilloid); TRPM; TRPA and TRPCs have proven excellent for studying

    Transient receptor potential ankyrin 1 (TRPA1) is activated by noxious cold (<17°C) and contributes to cold and mechanical hypersensitivity after inflammation and nerve injury: Yun Sook Kim, PhD, Hoon Kap Jung, DDS, Tae Kyung Kwon, DDS, Chin Soo Kim, DDS, PhD, Jin Hyun Cho, DDS, PhD, Dong Kuk Ahn, DDS, PhD, Yong Chul Bae, DDS, PhD.Expression of Transient Receptor Potential Ankyrin in Human Dental Pulp. Journal of Endodontics. Available online 8 June 2012.

    Highlights: TRPA1 was expressed in a large number of axons branching extensively in the peripheral pulp and in a few axons within the nerve bundles in the core of the coronal pulp and in the radicular pulp. Under electron microscopy, TRPA1 immunoreactivity was typically localized near the plasma membrane of unmyelinated axons in the peripheral pulp, suggesting that in these axons it may act as a functional receptor. The proportion of axons expressing TRPA1 in neurofilament 200–positive axons significantly increased in the painful pulp compared with the normal pulp. TRPA1 was also densely expressed in the processes and the cell body of odontoblasts. A large number of axons coexpressed TRPA1 and Nav1.8.

    Images: Immunofluorescent staining for (A) TRPA1 in the human normal dental pulp is completely abolished by (B) preadsorption with a control peptide, proving the specificity of the TRPA1 antiserum (×200, scale bars = 50 µm).
    Related Reagents:
    All TRP Antibodies                       
    Pain and Inflammation Research Antibodies                     
    Neurotransmission -Neurotransmission Research Antibody Categories
    Primary Neurons and Astrocytes-Primary human, rat and mouse neurons and astrocytes

    Monday, June 18, 2012

    Stem Cell Differentiation Flow Charts

    Installment #1: Mesoderm Derived Stem Cells

    We would like to thank our partners and customers for their input on how to best add value to our Stem Cell Research Reagents. Based on this input, we are in the process of building a new website focused on providing stem cell technical content. Better content should help catalyze better research and drug discovery related decisions.

    Here're several stem cell differentiation flow charts that map the genesis of pluripotent and multipotent cells and identify the cell types that result from their differentiation.

    This chart was generated using input and publications generously provided by Dr. Henry Young, Tenured Full Professor at Mercer University School of Medicine.

    I anticipate the launch date for our Stem Cell Related website to occur in early Q3 2012.

    In the meantime, we will posting relevant data, publications and other information here. This will include protocols and related growth factors for generating specific cell types like dopaminergic neurons from neural progenitors. Stay tuned.

    Wednesday, June 13, 2012

    Stress, Depression and Alpha2delta Ligands

    Implications for treating Stress Related Depression.

    Stress related disorders like Post Traumatic Stress Syndrome (PTSD), Major Depressive Disorder (MDD) and Generalized Anxiety Disorders (GAD) dysregulate neurogenesis. This disregulation can lead to disorders like chronic depression. Indeed, stress hurts.

    In this study (which includes use of our Neural Progenitor Marker-Nestin), researchers show for the first time that the alpha2delta (α2δ) ligands gabapentin [1- (aminomethyl)cyclohexaneacetic acid; GBP] and pregabalin [S-[+]-3-isobutylGABA or (S)-3- (aminomethyl)-5-methylhexanoic acid; PGB] can produce a concentration-dependent increase in the number of newborn mature and immature neurons generated in vitro from adult hippocampal neural progenitor cells (NPC), and, in parallel, a decrease in the number of undifferentiated precursor cells. These effects were confirmed in vivo, since a significantly increased number of adult generated neurons was observed in the hippocampal region of mice chronically treated with PGB [10 mg/kg, i.p., 21 days] compared to vehicle-treated mice. Moreover, we demonstrated that PGB administration prevented the appearance of depression-like behaviours induced by chronic restraint stress and, in parallel, promoted hippocampal neurogenesis in adult stressed mice. Finally, we provided data suggesting the potential involvement of the α2δ1 subunit and NF-κB signaling pathway in the drug-mediated proneurogenic effects. The new pharmacological activities of α2δ ligands may help explaining their therapeutic activity as add-on therapy in major depression and on depressive symptoms in posttraumatic stress disorder and generalized anxiety disorders. Furthermore these data contribute to the identification of novel molecular pathways which may represent potential targets for pharmacological modulation in depression: Maria Maddalena Valente, Valeria Bortolotto, Bruna Cuccurazzu, Federica Ubezio, Vasco Meneghini, Maria Teresa Francese, Pier Luigi Canonico, Mariagrazia Grilli.Alpha2delta ligands act as positive modulators of adult hippocampal neurogenesis andprevent depressive-like behavior induced by chronic restraint stress. Molecular Pharmacology Fast Forward Published on May 9, 2012 as doi:10.1124/mol.112.077636.

    Images: Effect of α2δ ligands on neuronal differentiation and proliferation of hippocampus-derived neural progenitor cells. (A) Representative fluorescence microscopy image of a hippocampal neurosphere immunolabelled for nestin (green) and SRY-related HMG-box gene 2 (Sox-2) (red), markers of undifferentiated NPC. Magnification = X600. Scale bar = 56 μm. (B) After 24 h in absence of growth factors, hippocampal Neural Progenitor Cells (NPC) differentiated giving rise to four different cell populations identified by double Microtubule Associated Protein-2 (MAP-2) and nestin immunolabelling: MAP-2+/nestin- mature neurons, MAP-2+/nestin+, MAP-2-/nestin+ and MAP-2-/nestin- cells. Data are expressed as mean ± S.D. of n=9 experiments, run in triplicates. Gabapentin (GBP) and pregabalin (PGB) promote neuronal differentiation of adult hippocampal NPC. GBP (C-F) and PGB (G-J) significantly increased, in a concentrationdependent manner, the percentage of MAP-2+/nestin- (C, G) and MAP-2+/nestin+ (D, H) cells and decreased the percentage of MAP-2-/nestin- cells (F, J), with no effect on MAP-2-/nestin+ cells (E, I). Data are expressed as mean ± S.D. of n = 3 experiments, run in triplicates. *, p < 0.05; **, p < 0.01; ***, p < 0.001 vs vehicle (Student’s t-test). (K-M) Representative fluorescence microscopy images of MAP-2 immunolabelling (green) in cells derived from hippocampal NPC after 24 h treatment with vehicle (K), 1 nM GBP (L) and 1 nM PGB (M). Nuclei are stained with Draq5(blue). Magnification = X400. Scale bar = 75 μm. (N) Adult hippocampal NPC were treated with vehicle or 1 nM PGB for 6, 24, 48, 72, 96 h and proliferation rate was assessed. PGB had no effect on NPC proliferation, when compared to vehicle. Data, expressed as counts per second (CPS), represent the mean ± S.D. of experiments run in triplicates.

    This study is good news. Results demonstrate the new pharmacological activity of α2δ ligands may

    potentially explain their efficacy as add-on therapy in MDD, as well as on depressive symptoms
    in PTSD and GAD. This knowledge will help the discovery of refined therapies for these debilating disorders.

    Saturday, June 09, 2012

    Serotonin and Pain

    Brainstem facilitations and descending serotonergic controls contribute to visceral nociception but not pregabalin analgesia in rats.

    Pregabalin is used to treat Neuropathic Pain. Here the authors show state-dependent pregabalin analgesia in neuropathy does not apply to visceral pain. Our rabbit polyclonal Mu Opioid Receptor Antibody is used to demonstrate intra-RVM Derm-SAP locally ablates a substantial proportion of MOR and serotonergic cells: Shafaq Sikandar, Kirsty Bannister, Anthony H. Dickenson. Brainstem facilitations and descending serotonergic controls contribute to visceral nociception but not pregabalin analgesia in rats. Neuroscience Letters. Volume 519, Issue 1, 21 June 2012, Pages 31–36.... For MOR staining, a rabbit polycolonal μ-opioid receptor primary antibody (1:10,000 TTBS; Neuromics, MN, USA, RA10104)...

    These findings are important for understanding the limitations of Preglabin as an analgesic for viceral pain. Check out all our Opioid Publications.