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人胚胎间充质干细胞参与新生小鼠肾小管细胞的分化。

Human embryonic mesenchymal stem cells participate in differentiation of renal tubular cells in newborn mice.

作者信息

Yuan Li, Liu Hou-Qi, Wu Min-Juan

机构信息

Department of Nephrology, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, P.R. China.

Department of Histology and Embryology, Research Center of Developmental Biology, Second Military Medical University, Shanghai 200433, P.R. China.

出版信息

Exp Ther Med. 2016 Aug;12(2):641-648. doi: 10.3892/etm.2016.3383. Epub 2016 May 24.

Abstract

Stem cells are used with increasing success in the treatment of renal tubular injury. However, whether mesenchymal stem cells (MSC) differentiate into renal tubular epithelial cells remains controversial. The aims of the present study were to observe the localization of human embryonic MSCs (hMSCs) in the kidneys of newborn mice, and to investigate hMSC differentiation into tubular epithelium. Primary culture hMSCs were derived from 4-7-week-old embryos and labeled with the cell membrane fluorescent dye PKH-26. The degree of apoptosis, cell growth, differentiation and localization of hMSCs with and without this label were then determined using immunohistochemical methods and flow cytometry. hMSCs and PKH26-labeled hMSCs were revealed to differentiate into chondrocytes and adipocytes, and were demonstrated to have similar proliferative capability. In the two cell types, the antigens CD34 and CD45, indicative of hematopoietic lineages, were not expressed; however, the expression of the mesenchymal markers CD29 and CD90 in MSCs, was significantly increased. During a 4-week culture period, laser confocal microscopy revealed that PKH26-labeled hMSCs in the kidneys of newborn mice gradually dispersed. Two weeks after the injection of the PKH26-labeled cells, the percentage of PKH26-labeled hMSCs localized to the renal tubules was 10±2.1%. In conclusion, PKH26 labeling has no effect on hMSC differentiation, proliferation and mesenchymal cell surface features, and hMSCs injected into the kidneys of newborn mice may transform to renal tubule epithelium.

摘要

干细胞在肾小管损伤治疗中的应用越来越成功。然而,间充质干细胞(MSC)是否分化为肾小管上皮细胞仍存在争议。本研究的目的是观察人胚胎间充质干细胞(hMSC)在新生小鼠肾脏中的定位,并研究hMSC向肾小管上皮细胞的分化情况。原代培养的hMSC来源于4 - 7周龄的胚胎,并用细胞膜荧光染料PKH - 26进行标记。然后使用免疫组织化学方法和流式细胞术确定有或无该标记的hMSC的凋亡程度、细胞生长、分化和定位情况。结果显示,hMSC和PKH26标记的hMSC均可分化为软骨细胞和脂肪细胞,且具有相似的增殖能力。在这两种细胞类型中,未表达指示造血谱系的抗原CD34和CD45;然而,间充质标记物CD29和CD90在MSC中的表达显著增加。在为期4周的培养期内,激光共聚焦显微镜观察显示,新生小鼠肾脏中PKH26标记的hMSC逐渐分散。注射PKH26标记细胞两周后,定位于肾小管的PKH26标记的hMSC百分比为10±2.1%。总之,PKH26标记对hMSC的分化、增殖和间充质细胞表面特征无影响,注入新生小鼠肾脏的hMSC可能转化为肾小管上皮细胞。

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