A Monolayer Culture Method for Neural Induction of hPSCs
Multipotent neural progenitor cells (NPCs) generate the major cell types of the central nervous system (CNS): neurons, astrocytes and oligodendrocytes. Human pluripotent stem cells (hPSCs), including embryonic stem (ES) cells and induced pluripotent stem (iPS) cells, can be directed to differentiate into NPCs using a variety of methods. This process, known as “neural induction”, must be efficient and reliable, in order to generate high-quality NPCs for downstream applications. hPSC-derived NPCs are used extensively for studying human nervous system development, modeling neurological disorders and screening potential therapeutic molecules. For a number of years, the predominant methods for neural induction of human ES and iPS cells have involved either co-culture with a stromal cell line, or embryoid body (EB) formation. More recently, studies have shown that neural induction from human ES and iPS cells can also be achieved in a monolayer culture system, wherein human ES and iPS cells are plated onto a defined matrix, and exposed to inductive actors.1,2 STEMdiff™ Neural Induction Medium is a defined, serum-free medium for the generation of NPCs from hPSCs. The standard protocol for this product involves EB formation and selection of neural rosettes, to obtain highly enriched cultures of CNS-type NPCs in 12 days (Figures 1A, 2A). In this technical bulletin, we describe a method for neural induction using STEMdiff™ Neural Induction Medium in a monolayer culture system (Figures 1B, 2B). Using this protocol, CNS-type NPCs can be generated from human ES or iPS cells in six to nine days.