The derived NSC are characterized by immunofluorescence with antibodies specific to nestin and SOX2. The cell population is highly pure: >95% of cells express Nestin and >95% of cells are SOX2 positive. H9-NSC are cryopreserved at P0 and delivered frozen. Each vial contains >1 x 106 cells in 1 ml volume. Cells are negative for mycoplasma, bacteria, yeast and fungi. After reviving, NSC can be maintained in HPSC Neural Induction Medium as an adherent culture. NSC are multipotent and able to differentiate into various neuronal and glial subtypes. Specific patterning cues, such as SHH, retinoic acid and FGF8, can be added after reviving to direct the cells to different neural lineages. To differentiate the H9-NSC, medium containing specific growth factors should be used (not provided).
Recommended Medium: It is recommended to use the provided Neural Induction Medium (PSCNIM, Cat. no. SC5931) for plating H9-NSC and expanding them in the short term. Adding ROCK inhibitor Y-27632 in the first 24 hours after reviving improves cell viability and attachment in adherent cultures.
Product Use: H9-NSC are for research use only. It is not approved for human or animal use, or for application in in vitro diagnostic procedures.
Storage: Upon receiving, directly and immediately transfer Cat. no. 1660 from dry ice to liquid nitrogen and keep the cells in liquid nitrogen until they are needed for experiments. Store Cat. no. 5931 at +4°C and Cat. no. 5982 at -20°C.
Shipping: Cat. no. 1660 and Cat. no. 5982 are shipped on dry ice. Cat. no. 5931 is shipped at room temperature.
References: [1] Chambers SM, Fasano CA, Papapetrou EP, Tomishima M, Sadelain M, Studer L. (2009) “Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling.” Nat Biotechnol. 27(3): 275-280.
[2] Li W, Sun W, Zhang Y, Wei W, Ambasudhan R, Xia P, Talantova M, Lin T, Kim J, Wang X, Kim W, Lipton SA, Zhang K, Ding S. (2011) “Rapid induction and long-term self-renewal of primitive neural precursors from human embryonic stem cells by small molecule inhibitors.” PNAS. 108(20): 8299-8304.