Wernig Marius, Tucker Kerry Lee, Gornik Volker, Schneiders Angelika, Buschwald Rachel, Wiestler Otmar D, Barde Yves-Alain, Brüstle Oliver
Institute of Reconstructive Neurobiology and Department of Neuropathology, University of Bonn Medical Center, Bonn, Germany.
J Neurosci Res. 2002 Sep 15;69(6):918-24. doi: 10.1002/jnr.10395.
Pluripotency and the capacity for continuous self-renewal make embryonic stem (ES) cells an attractive donor source for cell-replacement strategies. A key prerequisite for a therapeutic application of ES cells is the generation of defined somatic cell populations. Here we demonstrate that a targeted insertion of the EGFP gene into the tau locus permits efficient fluorescence-activated cell sorting (FACS)-based lineage selection of ES cell-derived neurons. After in vitro differentiation of heterozygous tau EGFP ES cells into multipotent neural precursors, EGFP is selectively induced in postmitotic neurons of various neurotransmitter phenotypes. By using FACS, ES cell-derived neurons can be enriched to purities of more than 90%. Because neuron-specific EGFP fluorescence is also observed upon transplantation of ES cell-derived neural precursors, the tau EGFP mutant represents a useful tool for the in vivo analysis of grafted ES cell-derived neurons.
多能性和持续自我更新的能力使胚胎干细胞（ES细胞）成为细胞替代策略中极具吸引力的供体来源。ES细胞治疗应用的一个关键前提是生成特定的体细胞群体。在此，我们证明将EGFP基因靶向插入tau基因座可实现基于高效荧光激活细胞分选（FACS）的ES细胞衍生神经元的谱系选择。杂合tau EGFP ES细胞在体外分化为多能神经前体细胞后，EGFP在各种神经递质表型的有丝分裂后神经元中被选择性诱导。通过使用FACS，ES细胞衍生的神经元可富集至纯度超过90%。由于在移植ES细胞衍生的神经前体细胞后也观察到神经元特异性EGFP荧光，tau EGFP突变体是体内分析移植的ES细胞衍生神经元的有用工具。
