Рагунепроцел

Rec. INN (наименование, зарегистрированное ВОЗ)

allogenic dopaminergic neural progenitor cells derived from human induced pluripotent stem cells (iPSC). The iPSC cell bank was established from peripheral blood mononuclear cells (PBMCs) isolated from a healthy human donor by episomal reprogramming with plasmid vectors expressing octamer-binding transcription factor 3/4 (OCT-3/4), SOX2, Krueppel-like factor 4 (KLF4), L-Myc, LIN28,tumor suppressor p53 (Tp53), and Epstein-Barr nuclear antigen 1 (EBNA1). The cells were initially cultivated with feeder-free stem cell culture media with interleukin-6 (IL-6), stem cell factor, thrombopoietin, Flt-3 ligand, IL-3, and granulocyte colony- stimulating factor. Following induction, the cells were seeded on plates coated with a truncated form of laminin in the same media. Clones were chosen on the best efficiency for subsequent dopaminergic differentiation.
The iPSCs were differentiated into dopaminergic progenitor cells in media containing a SMAD inhibitor, fibroblast growth factor 8 (FGF8), a Wnt inhibitor and Sonic Hedgehog (SHH).
CORIN (atrial natriuretic peptide-converting enzyme)-positive cells were then isolated by immunoselection, and further cultured under conditions that favour aggregation to promote progenitor spheres. The final cell population expresses the neuron-specific markers forkhead box protein A2 (FoxA2) and neuron-specific class III beta-tubulin (>90%) and are negative for the undifferentiated iPSC markers OCT 3/4 and alkaline phosphatase, tissue-nonspecific isozyme (TRA-2-49). Early neural progenitor cells are not present (no detectable levels of SOX1+ and PAX6+ double positive cells).

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