"Shenandoah Biotechnology"

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Cytokines for expansion and cell culture of human hematopoietic stem cells. Hematopoietic stem cells are self renewing pluripotent cells often characterised by the expression of CD34. Studies have shown that decreased expression of CD34 tends to correlate with decreased self renewal properties and pluripotent potential. Culturing CD34+ cells in a cytokine mixture of TPO, SCF, and FLT-3 Ligand for short time periods allows CD34+ cells to proliferate without decreasing CD34 expression, thus retaining their stem cell properties. The combination of these cytokines allows for the ex vivo expansion of the most immature hematopoietic stem cells in cord blood or bone marrow preparations, but can be supplemented with other cytokines to promote specific lineages. For example, GM-CSF and/or G-CSF can be added to promote monocytes and granulocytes while EPO can be added to promote erthroid cells.

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Interferon gamma (IFN-γ) is a type II interferon that is critical during adaptive and innate immune responses to infection. IFN-γ is produced by T cells and natural killer cells following antigen-specific activation. IFN-γ binds IFN-γ receptors (IFN-γ R1 and IFN-γ R2), which are expressed on most immune cells, to activate the JAK-STAT pathway. IFN-γ-induced signaling increases the expression of class 1 major histocompatibility complex (MHC) molecules. Human IFN-γ is not cross-reactive with mouse IFN-γ.

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Neuregulin 1-beta (NRG1-β) is one of many alternatively-spliced isoforms of the NRG1 gene and contains a soluble EGF-like domain. The EGF-like domain of NRG1-β signals through the ErbB2, ErbB3, and ErbB4 receptor tyrosine kinases. NRG1-β is an important growth factor involved in neuroinflammation, nerve regeneration, and cardiovascular processes.

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Insulin-like growth factor I (IGF-I) is a growth factor that is produced by the liver. IGF-1 production is stimulated by growth hormone (GH). IGF-I binds the insulin-like growth factor 1 receptor (IGF1R) and the insulin receptor to stimulate systemic body growth. IGF-I is one of the most potent activators of the AKT signaling pathway, which stimulates cell proliferation and inhibits programmed cell death. Mature human IGF-I is 100% homologous to bovine and porcine IGF-1 proteins.

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Activin A is a member of the transforming growth factor beta (TGF-β) family of proteins and functions to stimulate follicle-stimulating hormone (FSH) secretion. Activins are produced in many tissue types including the skin, gonads, lungs, and pituitary gland. Activins interact with receptor type I and type II serine/threonine protein kinases, to activate SMAD signaling and regulate diverse cellular functions, such as cell proliferation, differentiation, wound healing, apoptosis, and metabolism. Activin A is a homodimer comprised of two activin βA chains. Cleavage of the N-terminal propeptide renders the Activin protein biologically active. Human Activin A shares 100% amino acid sequence identity with mouse, rat, porcine, bovine, and feline Activin A proteins.

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Tumor necrosis factor alpha (TNF-α) is an inflammatory cytokine secreted by macrophages, monocytes, neutrophils, T cells, and NK-cells following stimulation by bacterial lipopolysaccharide (LPS).

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Stromal cell-derived factor-1 beta (SDF-1 β), also called CXCL12b, is one of two SDF-1 splice variants made by a wide variety of cells upon stimulation by inflammatory cytokines such as TNF, IL-1, and LPS. SDF-1 β signals through the G protein-coupled receptor CXCR4 to recruit activated leukocytes.

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Tumor necrosis factor alpha (TNF-α) is an inflammatory cytokine secreted by macrophages, monocytes, neutrophils, T cells, and NK-cells following stimulation by bacterial lipopolysaccharide (LPS). TNF-α signal activation occurs through two receptors, TNFR1 and TNFR2. TNFR1 is expressed on most cell types, unlike TNFR2, which is expressed mainly on immune cells. TNF-α functions to stimulate phagocytosis in macrophages, chemoattract neutrophils, increase insulin resistance, and induce fever.

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Interleukin 4 (IL-4) is an immunomodulatory cytokine that functions to induce naïve helper T cells to differentiate into type 2 T helper (Th2) cells. Th2 cells subsequently produce more IL-4 in a positive feedback loop. IL-4 also promotes immunoglobulin IgG to IgE isotype switching on B cells. IL-4 binds the IL-4Rα receptor to activate STAT6 signaling.

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Galectin-1 belongs to the lectin family of carbohydrate binding proteins and binds glycans as both a monomer and a homodimer. Galectin-1 is produced in peripheral lymphoid organs and inflammatory sites. Galectin-1 plays important roles in acute and chronic inflammatory processes, cell growth, cell proliferation, and induces apoptosis of activated T cells. Galectin-1 also modulates cytokine secretion and inhibits pro-inflammatory cytokine production.

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Epidermal growth factor (EGF) is a growth factor that stimulates the proliferation, differentiation, and survival of epithelial and epidermal cells. EGF contains three intramolecular disulfide bonds and binds in high affinity to the epidermal growth factor receptor (EGFR).

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Basic fibroblast growth factor (FGF-basic), also known as FGF-2, is expressed by endothelial cells and is a mediator of angiogenesis. FGF-basic also has cardioprotective functions during heart injury. FGF-basic is a critical component for embryonic stem cell culture systems and is necessary for maintaining cells in an undifferentiated state. Recombinant FGF-basic 154 is the full length FGF-basic protein encoded by the human FGF-2 gene. FGF-basic 154 is the most popular tissue culture product at Shenandoah Biotechnology, Inc. There are no detectable differences in biological activity between FGF-basic 154 and the truncated FGF-basic 147 proteins.

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Epstein-Barr virus induced gene-3 (EBI3) is a secreted glycoprotein belonging to the hematopoietin receptor family related to the p40 subunit of interleukin 12 (IL-12). EBI3 expression is induced in B-lymphocytes in response to Epstein-Barr virus infection. EBI3 forms heterodimers with p28 to form interleukin 27 (IL-27), and with p35 to form interleukin 35 (IL-35). Both IL-27 and IL-35 have anti-inflammatory and regulatory activity.

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Growth hormone is an important mitogenic growth factor that is synthesised, stored, and secreted by somatotropic cells of the anterior pituitary gland. They stimulates growth, cell reproduction, and cell regeneration.

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Cytokines for polarizing T cells into T regulatory cells. T regulatory cells (Tregs) are a subset of T cells that suppress immune function. They are characterized by the expression of CD4, CD25 and Foxp3 and are known to produce immunosuppressive cytokines TGF-β 1 and IL-10. Tregs can be cultured from naive CD4+ T cells by culturing them in the presence of IL-2 and TGF-β 1 with CD3 and CD28 stimulation. In some cases cells are also cultured with neutralizing antibodies to IL-4, IFNγ and IL-12 and/or retinoic acid.

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Procalcitonin is a precursor of the peptide hormone calcitonin. Procalcitonin is mainly produced by the neuroendocrine cells of the lung and thyroid gland, but may be secreted ubiquitously during inflammation or infection. Under normal expression conditions, procalcitonin is immediately cleaved into three specific fragments: calcitonin, katacalcin, and an N terminal residue. Levels of unprocessed procalcitonin are significantly increased after bacterial infection, inflammation, trauma, or shock.