Hwang Ji Hye, Yuk Soon Hong, Lee Jin Ho, Lyoo Won Suk, Ghil Sung-Ho, Lee Sang Sub, Khang In Gu, Paik Soon Young, Lee Ji Youl
Department of Microbiology, The Catholic University of Korea, Seoul 137-701, Korea.
Mol Cells. 2004 Feb 29;17(1):57-61.
We investigated whether stem cells (MDSC) from primary cultures of rat skeletal muscle can differentiate into the smooth muscle lineage in response to vascular endothelial growth factor (VEGF) and coculture with bladder smooth muscle cells. The MDSC were isolated from gastrocnemius muscle biopsies of normal 3-6 week-old Sprague-Dawley rats and purified by the preplate technique. Cells that took approximately 6 days to adhere to the collagen-coated flasks were termed late preplate (LP) cells, and were used in all the experiments. The early plate (EP) cells (pp1-pp4) contained some myogenic cells but were mostly fibroblasts (< 15% desmin+ cells) whereas the LP cells (pp5-pp6) were highly purified muscle-derived cells (pp6) (> 90% desmin+ cells). The muscle-derived stem cells (LP cells) were CD34+ or Sca-1+, CD45- and desmin+ by immunohistochemical staining. After two days of co-culture with bladder smooth muscle cells, about 25% of the muscle-derived stem cells were positive for alpha-smooth muscle actin (alpha-SMA)+. RT-PCR for alpha-SMA was positive in the VEGF stimulated MDSC, but negative in the absence of VEGF. In conclusion, rat muscle-derived stem cells exhibited stem cell properties (CD34+ or Sca-1+), and were not of hematogeous (CD45-) but of myogenic origin (desmin+). RT-PCR of alpha-SMA was positive in the VEGF stimulated muscle-derived stem cells.
我们研究了来自大鼠骨骼肌原代培养物的干细胞（MDSC）是否能响应血管内皮生长因子（VEGF）并与膀胱平滑肌细胞共培养而分化为平滑肌谱系。MDSC从3 - 6周龄正常Sprague-Dawley大鼠的腓肠肌活检组织中分离出来，并通过预铺板技术进行纯化。大约需要6天时间附着在胶原包被培养瓶上的细胞被称为晚期预铺板（LP）细胞，并用于所有实验。早期预铺板（EP）细胞（pp1 - pp4）含有一些成肌细胞，但大多是成纤维细胞（<15%结蛋白阳性细胞），而LP细胞（pp5 - pp6）是高度纯化的肌肉来源细胞（pp6）（>90%结蛋白阳性细胞）。通过免疫组织化学染色，肌肉来源的干细胞（LP细胞）为CD34 +或Sca - 1 +、CD45 -和结蛋白阳性。与膀胱平滑肌细胞共培养两天后，约25%的肌肉来源干细胞α - 平滑肌肌动蛋白（α - SMA）呈阳性。VEGF刺激的MDSC中α - SMA的RT - PCR呈阳性，但在无VEGF时为阴性。总之，大鼠肌肉来源的干细胞表现出干细胞特性（CD34 +或Sca - 1 +），不是造血来源（CD45 -）而是成肌起源（结蛋白阳性）。VEGF刺激的肌肉来源干细胞中α - SMA的RT - PCR呈阳性。
