Differentiation of human urine-derived stem cells into interstitial cells of Cajal-like cells by exogenous gene modification: A preliminary study.
Differentiation of human urine-derived stem cells into interstitial cells of Cajal-like cells by exogenous gene modification: A preliminary study.
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通过外源基因修饰将人尿液干细胞分化为 Cajal 样细胞间质细胞：初步研究。
DOI：
10.1016/j.bbrc.2019.09.121
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Li Longkun
中科院分区：
文献类型：
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作者：
Sun Bishao;Dong Xingyou;Zhao Jiang;Yang Zhenxing;Zhang Yuanyuan;Li Longkun
人尿源性干细胞（human urine-derived stem cells，hUSCs）具有多向分化潜能，可分化为中胚层细胞系。Cajal样细胞间质细胞（ICC-LC）是膀胱自发收缩的起搏功能的关键。然而，hUSC产生ICC-LC的机制尚未阐明。在这项研究中，我们开发了一种将hUSCs定向分化为ICC-LC的策略。用编码c-Kit、干细胞因子（SCF）、超极化激活的环核苷酸门控钾通道4（HCN 4）和5-氮杂胞苷诱导的2（AZI 2）基因的慢病毒载体转染hUSCs，并将细胞在特定培养基中再培养7天。在转染后7天测定ICC-LC上表面标记物c-Kit的表达。成功扩增hUSCs并用四种慢病毒载体转染。用慢病毒-c-Kit、慢病毒-HCN 4和慢病毒-AZI 2转染的hUSCs在转染后7天显示出更高的c-Kit表达，但只有慢病毒-HCN 4转染的细胞在ICC-LC中显示出形态学改变。这些细胞还显示出可见的HCN电流幅度和密度。这种方法可能为膀胱活动不足的治疗提供一种新的策略。
Human urine-derived stem cells (hUSCs) show multipotential differentiation ability and can differentiate into mesodermal cell lineages. Interstitial cells of Cajal-like cells (ICC-LCs) are crucial for the pace-making function of spontaneous contraction in the bladder. However, the mechanisms by which hUSCs generate ICC-LCs have not been elucidated. In this study, we developed a strategy for directional differentiation of hUSCs into ICC-LCs. hUSCs were transfected with lentiviral vectors encodingc-Kit, stem cell factor (SCF), hyperpolarization activated cyclic nucleotide gated potassium channel 4 (HCN4), and 5-azacytidine induced 2 (AZI2) genes, and the cells were cultured for an additional 7 days in specific medium. The expression of the surface marker c-Kit on ICC-LCs was determined at 7 days after transfection. hUSCs were successfully expanded and transfected with the four lentiviral vectors. hUSCs transfected with lentiviral-c-Kit, lentiviral-HCN4, and lentiviral-AZI2 showed higher expression of c-Kit 7 days after transfection, but only the lentiviral-HCN4-transfected cells showed morphological alterations in ICC-LCs. These cells also displayed visible HCN current amplitude and density. This approach may provide a new strategy for the treatment of underactive bladder.