小鼠血管外膜成纤维细胞原代培养及生物学特性

Primary culture and biological characteristics of adventitial fibroblasts in mice

目的 建立小鼠胸主动脉外膜成纤维细胞（adventitial fibroblasts,AF）组织块贴壁法培养模型.方法 组织块贴壁法培养小鼠AF,并在光镜下观察细胞形态,免疫细胞荧光染色法鉴定细胞类型,绘制细胞生长曲线.结果 AF能够迅速从组织块长出并增殖,光镜下细胞呈梭形或多边形,免疫荧光染色结果显示细胞质中有波形蛋白相关抗原存在,CD31和α-平滑肌肌动蛋白染色为阴性,细胞在第2～7天进入指数生长期.结论 组织块贴壁法适用于小鼠AF的原代培养,为研究AF生物学功能提供了充足、可靠的靶细胞模型.

Objective To establish the model by using tissue block pasted culture of adventitial fibroblasts （AFs） in mice.Methods Tissue block pasted culture method was employed to culture AFs,whose morpholog was observed under microscopy,and the immunocytofluorescence technique was applied to identify the type of cells,thus allowing for depiction of the growth curve.Results The isolated AFs that rapidly grew and proliferated were spindle-or polygon-shaped,and indicated the presence of vimentin-associated antigen with immunocytofluorescence staining.The CD31-and α-smooth muscle actin-associated antigens were negatively stained.The exponential phase of growth spanned from day 2 to day 7 following passage.Conclusion Tissue block pasted technique is effective for culture of AFs in mice and offers ample and reliable cell models for further investigation in the biological functions of AFs.