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Nature子刊:科学家将干细胞诱导分化获得浦肯野细胞

摘要 : 国际上首次将人源的诱导性多功能干细胞(iPSCs)诱导分化成为成熟具有电生理活性的浦肯野细胞

 首都医科大学宣武医院细胞生物室陈志国教授课题组在国际上首次将人源的诱导性多功能干细胞(iPSCs)诱导分化成为成熟具有电生理活性的浦肯野细胞,研究论文发表在3月18日发行的《Scientific Reports》上。该研究成果突破性地为研究浦肯野细胞(Purkinje cell)的功能,揭示小脑共济失调类疾病的发病机制奠定了有力的基础。

浦肯野细胞位于小脑浦肯野细胞层,是人脑内最大的神经元之一,也是小脑中唯一的传出神经元,在人体运动协调中发挥重要作用。浦肯野细胞损伤是小脑共济失调的主要病因之一,而长期以来,由于缺乏相应的细胞模型进行深入研究,这类疾病至今发病原因不明,临床上无良好治疗方案。课题组尝试将患有遗传性共济失调病人的皮肤成纤维细胞重新编程为诱导性多功能干细胞,并将其进一步诱导分化成为浦肯野前体细胞,通过共培养技术获得了成熟的人浦肯野细胞。

该研究成果为研究浦肯野细胞的功能,揭示小脑共济失调类疾病的发病机制奠定了有力的基础。同时该研究也证实,人浦肯野前体细胞移植到小鼠小脑后能够存活并继续分化成浦肯野细胞,因此该成果也为浦肯野细胞损伤和退行性疾病的细胞移植治疗带来了希望。

课题组专家表示,下一步研究中,将利用这一细胞资源深入研究此类遗传性共济失调及其相关疾病的发病机制,并在细胞水平进行药物筛选实验。

Differentiation of human induced pluripotent stem cells to mature functional Purkinje neurons

It remains a challenge to differentiate human induced pluripotent stem cells (iPSCs) or embryonic stem (ES) cells to Purkinje cells. In this study, we derived iPSCs from human fibroblasts and directed the specification of iPSCs first to Purkinje progenitors, by adding Fgf2 and insulin to the embryoid bodies (EBs) in a time-sensitive manner, which activates the endogenous production of Wnt1 and Fgf8 from EBs that further patterned the cells towards a midbrain-hindbrain-boundary tissue identity. Neph3-positive human Purkinje progenitors were sorted out by using flow cytometry and cultured either alone or with granule cell precursors, in a 2-dimensional or 3-dimensional environment. However, Purkinje progenitors failed to mature further under above conditions. By co-culturing human Purkinje progenitors with rat cerebellar slices, we observed mature Purkinje-like cells with right morphology and marker expression patterns, which yet showed no appropriate membrane properties. Co-culture with human fetal cerebellar slices drove the progenitors to not only morphologically correct but also electrophysiologically functional Purkinje neurons. Neph3-posotive human cells could also survive transplantation into the cerebellum of newborn immunodeficient mice and differentiate to L7- and Calbindin-positive neurons. Obtaining mature human Purkinje cells in vitro has significant implications in studying the mechanisms of spinocerebellar ataxias and other cerebellar diseases.

来源: Scientific Reports 浏览次数:12

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