Objective To culture C57BL/6 mouse pancreatic stellate cells (PSCs) with a modified method,and to establish a simple cultivation method with a high yield of PSCs.Methods Pancreatic tissues in C57BL/6 mice were collected,washed by fetal bovine serum,minced into 0.5 ～ 1 mm3,and cultured in sterile culture flasks.After 4days,primary PSCs were collected and passed on.The changes in intracellular lipid droplets were identified by oil red O staining;α-SMA,desmin expression was detected by immunocytochemical or immunocytofluorescent staining,and Western blot.Results Outgrowth cells in the fourth day were oval or star with abundant lipid droplets and fewer cells expressed c-SMA and desmin protein.PSCs turned into bigger,more pseudopodial ones with decreasing lipid droplets and significantly increasing expression of o-SMA and desmin protein after the passage.Protein expression of α-SMA in primary and passage PSCs were 0.653 ± 0.071,2.290 ± 0.055,respectively;and the expression in passage PSCs was significantly higher than that in primary PSCs (P ＜0.05),and the number of cells expressing desmin protein was significantly increased.Conclusions Modified tissue culture is a high yield and high purity method for both quiescent and activated PSCs,which can meet the need of in-vitro studies.

目的:建立一种简单的高效分离、培养、纯化和鉴定ICR小鼠骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSCs)的方法,初步研究其相关生物学特性.方法:全骨髓贴壁法分离培养BMSCs,用倒置显微镜观察细胞的形态和生长情况;用流式细胞仪鉴定细胞表面标记物;诱导细胞定向分化为成脂细胞、成骨细胞,以检测细胞多向诱导分化能力.结果:利用该方法纯化的BMSCs形态均一,多为梭形.第3代BMSCs进行流式检测CD29、CD44均呈阳性表达,表达率分别为97％、90％;CD34、CD45呈阴性表达;且能够成功诱导分化成成骨细胞、成脂细胞.结论:利用全骨髓贴壁法可以高效分离、纯化ICR小鼠的BMSCs,生物学特性稳定.