Basic fibroblast growth factor (bfgf)

HNPCs were prepared from the central nervous system of first-trimester human fetuses, as described previously [28 (link)]. Medium, referred to as N2A medium, was changed 3 times a week, and the growth factors EGF and bFGF (both at 20 ng/ml; Abcys, Eurobio, France) were added to maintain undifferentiated cells. The population of undifferentiated cells is composed exclusively by hNPCs. The D0 cell populations are composed mainly by hNPCs and in a smaller proportion of neurons and astrocytes whereas the D5 cells are mainly composed of neurons and astrocytes.

BM-hMSCs were cultured as previously described [28 (link)]. Briefly, cells were seeded on a cell culture-treated surface (Corning, New York, USA) in the presence of Minimum Essential Medium (MEMα) manufactured under GMP conditions (Macopharma) and supplemented with either MSC-qualified FBS (Gibco, Life Technologies, Carlsbad, USA) with 1 ng/mL bFGF (Eurobio, Montpellier, France) or hPL or PR-hPL. Heparin (Biochrom, VWR, Radnor, USA) at 2 IU/mL was added to hPL- and PR-hPL-containing media to avoid gelation of the medium. 100 U/mL penicillin G / 0.1 mg/mL streptomycin sulfate (Lonza, Basel, Switzerland) was added under all conditions, and the media were renewed twice a week. Cell cultures were maintained in a humidified atmosphere containing 5% CO₂. All experiments were performed between P1 and P4.