摘要
目的建立肾脏成肌纤维细胞的培养方法，为肾脏纤维化的发病机制和防治措施体外研究提供细胞技术平台。方法采用肾皮质组织块接种培养法，培养的细胞通过相差显微镜观察形态、电镜下观察细胞器，细胞免疫化学染色检测细胞表型标记蛋白等进行鉴定。同时检测细胞对不同细胞因子的生长反应。结果培养的细胞呈梭形，单个核，多突起，表达波形蛋白（vimentin）和平滑肌肌动蛋白（α，SMA），不表达主要存在于血管内皮细胞的抗原上皮氨基肽酶P（EAP），不表达角蛋白（cytokerafin）和结蛋白（desmin）。细胞可以传代至5代，并且保持成肌纤维细胞表型。转化生长因子β1、结缔组织生长因子刺激细胞增殖，而γ-干扰素对细胞无增殖效应。结论原代培养细胞为成肌纤维细胞并且可以传代，具备对细胞因子的生长调节反应，为后续实验提供了基础。
Objective To establish a method culturing myofibroblasts primarily from cortex of rat kidney. Methods Explanting the pieces of cortex of rat kidney into plastic flask, then cells grew out from the tissue pieces. Cells were identified by observation with inverted phase contrast microscope and electron microscope, as well as by phenotypic protein examination via immunochemistry staining and Western blot analysis. Results The cells showed shuttle shape with single nuclear in each one. Expression of vimentin and α-SMA was positive over all of the cells, which confirmed the myofibroblast phenotype. In contrast, expression of epithelial aminopeptidase P, cytokeratin, desmin, which were known as the markers for both endothelial cells and epithelial cells, was negative in these cells. Cultured cells could be propagated to 5 passage and remained the phenotype of myofibroblsts. Connective tissue growth factor （CTGF） and transforming growth factor β1 （TGF-β1） could stimulate cell proliferation, but interferon-gamma had no effect on these cells. Conclusion Cultured myofibroblasts from the cortex of rat kidney can be propagated and have proliferative response to CTGF and TGF-β1.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2007年第1期44-48,共5页
Chinese Journal of Nephrology
基金
国家自然科学基金（30570851）
教育部新世纪优秀人才基金（985-2-071-113） 感谢本研究所细胞研究室技术员唐嘉薇和刘颖在细胞培养和免疫化学方法方面给予的指导和帮助
关键词
肾皮质
成肌纤维细胞
原代培养
Cortex of kidney
Myofibroblasts
Primary culture
