Kim Mijeong, Habiba Ayman, Doherty Jason M, Mills Jason C, Mercer Robert W, Huettner James E
Department of Cell Biology and Physiology, Washington University Medical School, 660 South Euclid Avenue, St. Louis, MO 63110, USA.
Dev Biol. 2009 Apr 15;328(2):456-71. doi: 10.1016/j.ydbio.2009.02.001. Epub 2009 Feb 13.
Pluripotent mouse embryonic stem cells (ESCs) derived from the early blastocyst can differentiate in vitro into a variety of somatic cell types including lineages from all three embryonic germ layers. Protocols for ES cell neural differentiation typically involve induction by retinoic acid (RA), or by exposure to growth factors or medium conditioned by other cell types. A serum-free differentiation (SFD) medium completely lacking exogenous retinoids was devised that allows for efficient conversion of aggregated mouse ESCs into neural precursors and immature neurons. Neural cells produced in this medium express neuronal ion channels, establish polarity, and form functional excitatory and inhibitory synapses. Brief exposure to RA during the period of cell aggregation speeds neuronal maturation and suppresses cell proliferation. Differentiation without RA yields neurons and neural progenitors with apparent telencephalic identity, whereas cells differentiated with exposure to RA express markers of hindbrain and spinal cord. Transcriptional profiling indicates a substantial representation of transit amplifying neuroblasts in SFD cultures not exposed to RA.
源自早期囊胚的多能小鼠胚胎干细胞（ESCs）在体外可分化为多种体细胞类型，包括来自所有三个胚胎胚层的谱系。ES细胞神经分化方案通常涉及视黄酸（RA）诱导，或暴露于生长因子或其他细胞类型条件培养液中。设计了一种完全不含外源性类视黄醇的无血清分化（SFD）培养基，该培养基可使聚集的小鼠ESCs高效转化为神经前体和未成熟神经元。在此培养基中产生的神经细胞表达神经元离子通道，建立极性，并形成功能性兴奋性和抑制性突触。在细胞聚集期间短暂暴露于RA可加速神经元成熟并抑制细胞增殖。无RA分化产生具有明显端脑特征的神经元和神经祖细胞，而暴露于RA分化的细胞则表达后脑和脊髓的标志物。转录谱分析表明，在未暴露于RA的SFD培养物中，过渡放大神经母细胞大量存在。
