Hi All,

We are attemting to differentiate the ReNcell CX Human Neural Progenitor Cell Line (https://www.sigmaaldrich.com/GB/en/product/mm/scc007) into neurons and astrocytes, as described in the company's protocol (https://www.sigmaaldrich.com/GB/en/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/stem-cell-culture/rencell-human-neural-progenitors).

However, we are currently unable to induce differentiation as suggested, which is the removal of growth factors bFGF and EGF from the medium. The cells continue to proliferate to 100% confluency after 3-4 days, even when seeded at around 50% confluency, and don't display any morphological signs of differentiation during that time (up to 2 weeks). In some cases, I've attempted to seed at lower confluence, but the cells appear to display stress at this process. We have attempted this using Rencell Maintenance media and DMEM/F12 (+B27 and heparin) media, and coat flasks with 20 ug/ml Laminin.

We are currently attempting to push the differentiation using factors including BDNF, however given that I haven't seen this in any protocols for this cell line, I also wanted to see whether anyone had any other advice for differentiating the Rencell CX culture?

Any help/advice is greatly appreciated!