Human Neural Stem Cells from AcceGen are differentiated from a human induced pluripotent stem cell line (hiPSC), which is generated using mRNA reprogramming technology from Human Dermal Fibroblasts (HDF). The monolayer HDF-hiPSC are efficiently converted to neural epithelium using HPSC Neural Induction Medium, a serum-free medium for rapid and efficient neural induction of human pluripotent stem cells (hPSCs). Synergistic inhibition of glycogen synthase kinase 3 (GSK3) and transforming growth factor b (TGF-b) differentiates HDF-hiPSCs to homogenous neural stem cells (NSC) in the presence of human leukemia inhibitory factor (LIF) within 7 days. The derived NSC are characterized by immunofluorescence with antibodies specific to nestin and SOX2. The cell population is highly pure: >90% of cells express Nestin and >80% of cells are SOX2 positive. After reviving, NSC can be maintained in HPSC Neural Induction Medium as an adherent culture or form neural spheres in EGF and bFGF containing neural stem cell medium. The cells are able to form rosette and neural tube-like structures after being passaged and plated at high density on Matrigel coated culture vessels in neural stem medium containing EGF and bFGF. HiPSC-NSC can be further expanded for months as a monolayer on MatrigelTM coated culture vessels or in suspension as neural spheres when seeded on ultra-low binding culture vessels in neural stem cell medium. 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. HiPSC-NSC are cryopreserved at passage 0 and delivered frozen. Each vial contains >1 x 10^6 cells in 1 ml volume. Cells are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast and fungi.

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