Ma Lixiang, Liu Yan, Zhang Su-Chun
Department of Anatomy, Histology and Embryology, Shanghai Medical College, Fudan University, Shanghai, China.
Methods Mol Biol. 2011;767:411-8. doi: 10.1007/978-1-61779-201-4_30.
Midbrain dopaminergic (mDA) neurons play a critical role in regulating postural reflexes and movement as well as modulating psychological processes. Dysfunction or degeneration of mDA neurons is involved in a number of neurological disorders including Parkinson's disease. Availability of large quantities of human mDA neurons would greatly enhance our ability to reveal pathological processes underlying mDA neuron degeneration and to identify treatments for these neurological conditions. Human pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), provide an unlimited source for mDA neurons. Here we describe a chemically defined protocol for mDA neuron differentiation. PSCs are first converted to neuroepithelia in a chemically defined medium without any growth factors, followed by patterning the neuroepithelia to midbrain progenitors with fibroblast growth factor 8 (FGF8) and sonic hedgehog (SHH) and subsequent differentiating to functional mDA neurons. This protocol typically yields about half of the neuronal population being mDA neurons, determined by expression of mDA markers, electrophysiological recordings, and the ability to reverse functional deficit in a rat model of Parkinson's disease.
中脑多巴胺能（mDA）神经元在调节姿势反射和运动以及调节心理过程中起着关键作用。mDA神经元的功能障碍或退化与包括帕金森病在内的多种神经系统疾病有关。大量人类mDA神经元的可获得性将极大地增强我们揭示mDA神经元退化潜在病理过程以及确定这些神经系统疾病治疗方法的能力。人类多能干细胞（PSC），包括胚胎干细胞（ESC）和诱导多能干细胞（iPSC），为mDA神经元提供了无限的来源。在这里，我们描述了一种用于mDA神经元分化的化学成分明确的方案。PSC首先在不含任何生长因子的化学成分明确的培养基中转化为神经上皮，然后用成纤维细胞生长因子8（FGF8）和音猬因子（SHH）将神经上皮诱导为中脑祖细胞，随后分化为功能性mDA神经元。通过mDA标记物的表达、电生理记录以及在帕金森病大鼠模型中逆转功能缺陷的能力来确定，该方案通常产生约一半的神经元群体为mDA神经元。
