Derivation of homogeneous GABAergic neurons from mouse embryonic stem cells

• Published in: Experimental neurology Vol. 217; no. 2; pp. 407 - 416
• Main Authors: Chatzi, Christina, Scott, Roderick H., Pu, Jin, Lang, Bing, Nakamoto, Chizu, McCaig, Colin D., Shen, Sanbing
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.06.2009; Elsevier
• Subjects: Animals; Biological and medical sciences; Biomarkers - analysis; Biomarkers - metabolism; Cell Culture Techniques; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cells, Cultured; Central nervous system; Development. Senescence. Regeneration. Transplantation; Embryoid body; Embryonic stem cells; Embryonic Stem Cells - cytology; Embryonic Stem Cells - drug effects; Embryonic Stem Cells - physiology; Fundamental and applied biological sciences. Psychology; GABAergic neurons; gamma-Aminobutyric Acid - metabolism; Interneurons - cytology; Interneurons - metabolism; Medical sciences; Mice; Neurogenesis - drug effects; Neurogenesis - physiology; Neurology; Potassium Channels, Voltage-Gated - metabolism; Retinoic acid; Spheroids, Cellular; Stem Cell Transplantation - methods; Tretinoin - pharmacology; Vertebrates: nervous system and sense organs; Embryoid body; GABAergic neurons; Embryonic stem cells; Retinoic acid; Nervous system diseases; Stem cell; Rodentia; Retinoid; Vertebrata; Mammalia; Mouse; Animal; Gabaergic neuron
• ISSN: 0014-4886; 1090-2430; 1090-2430
• DOI: 10.1016/j.expneurol.2009.03.032

Embryonic stem cells (ESCs) promise an unlimited source of defined cells for cell transplantation therapy, while protocols for derivation of homogeneous populations of desirable cell types are yet to be developed and/or refined. Gamma aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the central nervous system, and disturbed GABAergic signaling is associated with a host of neurological conditions. We developed a simple ES cell differentiation protocol which led to the production of uniform GABAergic neurons in ∼ 2 weeks. The differentiation protocol involved treatment of embryoid bodies (EBs) with high concentrations (10 −5–10 − 4 M) of all-trans-retinoic acid (RA) for 3 days. After plating these EBs on attached dishes in neural supportive medium, 93–96% of the cells became GABA-positive neurons in 7–11 days. These cells also expressed immature neuronal markers with voltage-gated delayed rectifier potassium currents, suggesting that they were immature GABAergic neurons. The technology may have implications for modeling and treatment of GABAergic signaling-related diseases and injuries.