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Product Details:Tissue Human spinal nervePackage Size 0.5 million cells/vialPassage Number P1Shipped CryopreservedStorage Liquid nitrogenGrowth Properties AdherentMedia Schwann Growth medium (cat # SGM001).Cat# SC001Product Description:Schwann cells are neural crest derivatives that ensheathe and myelinate axons of peripheralnerves. Each Schwann cell wraps around the shaft of an individual peripheral axon, formingmyelin sheaths along segments of the axon. Schwann cells play important roles in thedevelopment, function, and regeneration of peripheral nerves. When an axon is dying, theSchwann cells surrounding it aid in its digestion, leaving an empty channel formed by successiveSchwann cells, through which a new axon may then grow from a severed end. The number ofSchwann cells in peripheral nerves is tightly regulated. Their proliferation in vitro can bestimulated by various growth factors including PDGF, FGF, neuregulin, and others. Schwanncells provide a relatively simple, well-defined, and accessible mammalian model for the study ofa number of developmental questions. It is also of particular clinical importance to understandthe biology of Schwann cells, not only in the context of neuropathies and nerve regeneration,but also because the cells or their precursors may be especially well suited for implants tofacilitate repair in the CNS.Human Schwann Cells (HSwC) from Alphabioregenare isolated from human spinalnerve. HSwC are cryopreserved at passage one and delivered frozen. Each vial contains >5 x10^5 cells in 1 ml volume. HSwC are characterized by immunofluorescence with antibodiesspecific to S100, GFAP, and CD90. HSwC are negative for HIV-1, HBV, HCV, mycoplasma,bacteria, yeast, and fungi. HSwC are guaranteed to further expand for 10 population doublingsin or Schwann Growth medium (cat # SGM001)
Live/proliferating cells:1. Upon arrival, incubate the flask containing cells and media in an incubator (37°C, 5% CO2) for3-5 hours to recover from transportation.2. Carefully place the vessel in a biosafety cabinet and spray the outer side of flask with 70%ethanol to disinfect.3. Let it air dry. Carefully open the vessel while keeping it in upright standing position.4. If the cells you received are suspension carefully transfer cell suspension is ready to be platedin the desired culture flask containing appropriate growth media. Incubate again the cultureflask in incubator (37°C, 5% CO2). Change media or subculture as needed.5. If the cells you received are adherent: Carefully aspirate the media and add fresh appropriategrowth media to flask and let it incubate overnight at 37°C, 5% CO2). Change media orsubculture as needed.Frozen Cells Protocol:1. Remove the cryovial from the liquid nitrogen storage tank.2. Thaw the cells quickly by placing the lower half of the vial into a 37°C water bath whileagitating gently, remove after 60 seconds. Keep the cap out of the water to avoidcontamination. There should still be a few ice crystals left after thawing (it is important not toover-thaw the cryovials as the presence of DMSO is toxic to the cells).3. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol.From this point onwards, all operations should be strictly carried out inside a laminar flow hoodin aseptic conditions.
4. Gently re-suspend the cells in the vial and transfer the cell suspension into a 15 mL sterileconical tube containing 5 mL of pre-warmed, complete media using a sterile transfer pipette.5. Centrifuge the cells at 1500 rpm for 3 minutes to pellet (the actual centrifuge duration andspeed may vary, depending on the cell type).6. Check the clarity of the supernatant to ensure that all cells are pulled down into the pellet.Aspirate out the supernatant without disturbing the cell pellet.7. Re-suspend the cell pellet in fresh, pre-warmed culture media and transfer the cells into aculture vessel. Gently rock the culture vessel to distribute the cells evenly. Table 1 providesguidelines to the volume of culture media needed for a range of culture vessels.8. Incubate the culture at 37°C, 5% CO2, or another recommended culture environmentspecified by the cell line datasheet. Incubate for at least 24 hours before processing the cells fordownstream experiments.Freezing Protocol:Always freeze down cells at a high concentration and at as low a passage number as possible.Ensure that the cells are at least 90% viable before freezing. Always use proper aseptictechnique and work in a laminar flow hood. Always wear personal protective equipment whenworking with liquid nitrogen.1. Harvest log phase cells (with > 90% viability): For adherent cells, gently detach the cells fromthe culture vessel to collect cells into a centrifuge tube following the Subculturing Protocol(step 1 to step 6). For suspension cells, harvest all cells into a centrifuge tube.2. Determine viable cell density and calculate the required volume of CryopreservationMedium needed (FM300). We recommend freezing cells at 1.0 to 2.5 x 106 cells/ml.
3. Centrifuge the cell suspension at 1500 rpm for 3 minutes.4. Aseptically, aspirate out the supernatant without disturbing the pellet.5. Re-suspend the cell pellet in Cryopreservation Medium at the appropriate cell density.6. Dispense the cell suspension into cryovials and freeze according to your laboratory standard(i.e. controlled rate freezing at approximately 1 oC decrease per minute).7. Transfer the frozen cells into liquid nitrogen storage (in the gas phase above the liquidnitrogen) for long-term storage.