Generation of dorsal spinal cord GABAergic neurons from mouse embryonic stem cells.

Developmental signaling molecules involved in dorsal patterning of the spinal cord have been identified in vivo; however, studies have not produced specific functional dorsal spinal cord neurons in vitro. We present here differentiation of R1 embryonic stem (ES) cells into GABAergic dorsal spinal cord neurons by sequential treatment with developmental signaling molecules. We found that retinoic acid, Bmp4 altered the specification of neural progenitors and instructed neural fate when applied at distinct stages of development. High concentration of retinoic acid initiated caudal patterning during early differentiation; Bmp4 induced dorsal development. The combination of retinoic acid and different concentration Bmp4 controlled the differing regional progenitors of spinal cord. Low-concentration Bmp4 and high concentration of retinoic acid-treated embryoid bodies resulted in the differentiation of GABAergic neurons. In summary, we demonstrate this simple treatment paradigm produced simple dorsal spinal cord neurons, which could be utilized for developmental and preclinical studies.