Generation of Schwann cell progenitors from murine ES cells

Abstract

Schwann cells (SCs) constitute the main glia population of the peripheral nervous system. Irreparable damages of Schwann cells are the cause of a broad spectrum of neurological diseases. In vitro differentiation of embryonic stem (ES) cells could represent an attractive strategy to derive SCs in large numbers in vitro. We have recently developed methods for the derivation of CNS glia from murine ES cells. Here we set out to explore whether BMP-induced promotion of neural crest differentiation can be exploited for the generation of ES cell-derived SCs. Following growth factor withdrawal, ES cell-derived neural precursors propagated in 10 ng/ml FGF2 and 5 ng/ml BMP2 generated 8±2.5% P0-positive putative SCs as compared to 1,5±0,5% P0-positive cells in non BMP-treated controls. RT-PCR, Western blot and immunofluorescence analyses showed that treatment with forskolin and neuregulin-I (NRG-I) further promotes expression of the SC marker Krox20, yielding up to 42±10% immunoreactive cells. Combined with immunopanning with an anti p75 antibody, enrichment of Krox20-positive cells could be further enhanced to 69+10% of the total population. Considering their capacity for the myelin formation in both the central and the peripheral nervous system, ES cell-derived SCs provide attractive perspectives for neural repair.