Rolletschek A, Chang H, Guan K, Czyz J, Meyer M, Wobus A M
'In vitro Differentiation' Group, Institute of Plant Genetics and Crop Plant Research, Corrensstrasse 3, D-06466, Gatersleben, Germany.
Mech Dev. 2001 Jul;105(1-2):93-104. doi: 10.1016/s0925-4773(01)00385-9.
Here, we describe the generation of viable and dopamine-producing neurons derived from pluripotent mouse embryonic stem cells. Neurotrophic factors in combination with survival-promoting factors, such as interleukin-1beta, glial cell line-derived neurotrophic factor, neurturin, transforming growth factor-beta(3) and dibutyryl-cyclic AMP, significantly enhanced Nurr1 and tyrosine hydroxylase (TH) mRNA levels, whereas En-1, mash-1 and dopamine-2-receptor mRNA levels were not upregulated. In parallel, mRNA levels of the anti-apoptotic gene bcl-2 were found to be upregulated at terminal stages. Double immunofluorescence analysis revealed increased numbers of TH- and dopamine transporter-, but not gamma-aminobutyric acid- and serotonin-positive neurons in relation to synaptophysin-labeled cells by survival-promoting factors. Moreover, high-performance liquid chromatography analysis showed detectable levels of intracellular dopamine. We conclude that survival-promoting factors enhance differentiation, survival and maintenance of dopaminergic neurons derived from embryonic stem cells.
在此，我们描述了从多能小鼠胚胎干细胞衍生出有活力且能产生多巴胺的神经元的过程。神经营养因子与促存活因子（如白细胞介素-1β、胶质细胞系源性神经营养因子、神经营养素、转化生长因子-β(3)和二丁酰环磷腺苷）联合使用，可显著提高Nurr1和酪氨酸羟化酶（TH）的mRNA水平，而En-1、mash-1和多巴胺-2-受体的mRNA水平未上调。同时，发现抗凋亡基因bcl-2的mRNA水平在终末阶段上调。双重免疫荧光分析显示，与突触素标记的细胞相比，促存活因子使TH和多巴胺转运体阳性神经元数量增加，但γ-氨基丁酸和5-羟色胺阳性神经元数量未增加。此外，高效液相色谱分析显示细胞内多巴胺水平可检测到。我们得出结论，促存活因子可增强源自胚胎干细胞的多巴胺能神经元的分化、存活和维持。
