成年鼠骨髓单个核细胞在体外培养中分化为平滑肌祖细胞.docVIP

【摘要】 　　目的：探讨大鼠骨髓单个核细胞在体外经条件培养基诱导定向分化成平滑肌祖细胞的可行性。方法：分离4～6周龄的大鼠胫骨、股骨，以4℃预冷的DMEM培养基冲出骨髓，密度梯度离心法分离骨髓单个核细胞，在血小板源生长因子-BB （ PDGF-BB）和碱性成纤维细胞生长因子 （ b-FGF）作用下，培养8～12d形成贴壁的梭形细胞。采用CD34与α-SMA免疫荧光染色进行鉴定，RT-PCR法测定其α-SMA mRNA表达情况。结果： 在PDGF-BB作用下，82%的骨髓单个核细胞α-SMA染色阳性，78%细胞CD34染色阳性，新鲜分离的骨髓单个核细胞不表达α-SMA mRNA，体外培养后表达α-SMA mRNA。结论：体外培养的骨髓单个核细胞能分化为平滑肌祖细胞，可作为研究平滑肌细胞分化和筛选抑制再狭窄药物的工具。 【关键词】 骨髓细胞；干细胞；肌细胞，平滑肌 　　Bone marrow mononuclear cells of adult rat to differentiate into smooth muscle progenitor cells in vitro/ZHANG Po, HUNAG Lan, SONG Ming|bao, ZHAO Xiao|Hui，CUI Bin, ZHOU Yin|pin, YIN Yang|guang, ZHU Guang|xu//Chinese Journal of Cardiovascular Rehabilitation Medicine,2007，16（2）：128 Abstract：Objective:To explore the potential of rat bone marrow mononuclear cells (MNCs) to differentiate into vascular smooth muscle progenitor cells.Methods:After the tibias and femurs were dissected from 4~6 week|old rats, the marrow was flushed out with ice|cold DMEM mediumThe MNCs were obtained with density gradient centrifugation and cultured in DMEM medium containing 20%FBS, 50ng/ml Platelet|derived growth factor-BB(PDGF-BB), 10ng/ml basic fibroblast growth factor(b-FDF).The cultured cells in vitro were identified with smooth muscle cell|specific α actin (α-SMA) and CD34 by immunofluorescent techniqueRT-PCR technique was used to identify the α-SMA mRNA level in fresh isolated marrow mononuclear cells and SPCs.Results:Smooth muscle|like cells outgrew from the culture of MNCs in presence of PDGF-BB and b-FGF.These cells were positive stain for α-SMA and CD34 on immunofluorescenceα-SMA mRNA was not expressed in fresh isolated MNCs but in the cells cultured for 8-12 days.Conclusion:Vascular smooth muscle cells can outgrow from bone MNCsThis model may be useful in study of smooth muscle cell differentiation and the origin of neo|intimal cells after vascular injury. Author′s　address：Cardiovascular Research Institute of PLA, Xinqiao Hospital of the Military Medical University, Chongqing 400037, C