Primary human NPCs were obtained from fetal brain tissue assumed to be derived from the dorsal telencephalon based on visual inspection at approximately 14 to 21 gestation weeks, as previously described (35 (link)–37 (link)). After single-cell dissociation, cells were initially cultured as neurospheres before plating on fibronectin-coated (F1141; Sigma-Aldrich) and poly-L-ornithine–coated (P3655; Sigma-Aldrich) plates, passaged 2 or 3 times, cryopreserved, and transferred to the University of North Carolina at Chapel Hill. NPC media (Neurobasal A, 10888-022; Life Technologies) was supplemented with 100 μg/mL primocin (ant-pm-2; InvivoGen), 10% BIT 9500 (09500; StemCell Technologies), 1% glutamax (100x, 35050061; Life Technologies), 1 μg/mL heparin (H3393-10KU; Sigma-Aldrich), 20 ng/mL epidermal growth factor/fibroblast growth factor (PHG0313/PHG0023; Life Technologies), 2 ng/mL leukemia inhibitory factor (PHC9481; Life Technologies), and 10 ng/mL platelet-derived growth factor (PHG1034; Life Technologies). We followed previously established protocols to maintain NPCs as proliferating neural progenitors and inhibit differentiation into neurons (36 (link)). No mycoplasma contamination was detected during regular preassay screens of cell culture media (30-1012K; ATCC).
Phg0313 phg0023
Manufactured by Thermo Fisher Scientific
The PHG0313/PHG0023 is a laboratory equipment product from Thermo Fisher Scientific. It is a pH meter designed for routine pH measurements in various laboratory applications. The device provides basic pH measurement functionality.
Automatically generated - may contain errors
Lab products found in correlation
Phc9481, by Thermo Fisher Scientific (2 mentions)
Phg1034, by Thermo Fisher Scientific (2 mentions)
F1141, by Merck Group (1 mentions)
P3655, by Merck Group (1 mentions)
Primocin ant pm 2, by InvivoGen (1 mentions)
Poly l ornithine, by Merck Group (1 mentions)
Neurobasal a, by Thermo Fisher Scientific (1 mentions)
Bit 9500, by STEMCELL (1 mentions)
2 protocols using phg0313 phg0023
1
Culturing Human Neural Progenitor Cells
2
Derivation of Human Neural Progenitor Cells
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Fetal brain tissue visually consistent with dorsal telencephalon morphology (flat and sheet-like) was collected at 14–21 gestation weeks from presumed neurotypical donors to derive primary human NPCs as previously described10 (link),11 (link),26 (link). To summarize, tissue was dissociated into single cells which were cultured as neurospheres before transfer to fibronectin (Sigma F1141) and Poly-L-Ornithine (Sigma P3655) coated plates. Neuronal differentiation was suppressed to maintain NPCs in a proliferative state by following previously established culture methods10 (link). After 2–3 passages, NPC lines were cryopreserved and transferred to UNC Chapel Hill. NPC media: Neurobasal A (Life Technologies 10888-022) supplemented with 100 μg ml−1 primocin (Invivogen ant-pm-2), 10% BIT 9500 (Stemcell Technologies 09500), 1% glutamax (100x; Life Technologies 35050061), 1 μg ml−1 heparin (Sigma-Aldrich H3393-10KU), 20 ng ml−1 EGF/FGF (Life Technologies PHG0313/PHG0023), 2 ng ml−1 LIF (Life Technologies PHC9481) and 10 ng ml−1 PDGF (Life Technologies PHG1034).
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