目的：研究成体小鼠心脏Sca-1＋细胞体外诱导分化的潜能。方法：利用免疫磁珠分选法分离成体小鼠心脏Sca-1＋细胞，通过BMP-2／FGF-4，TGF-β1及VEGF165等三种不同的体外诱导方法使其向心脏“三系”分化，并从细胞形态的变化、RT-PCR和免疫荧光染色等方面鉴定其分化潜能。结果：经BMP-2／FGF-4诱导，小鼠心脏Sca-1＋细胞发生形态改变，上调心肌细胞特征基因α-MHC、β-MHC、MLC-2a和MLC-2v的mRNA表达，同时表达心肌特征性蛋白cTNT和cMHC。经TGF-β1诱导，小鼠心脏Sca-1＋细胞亦发生形态改变，上调平滑肌特征基因α-SMA和Calponin的mRNA表达，同时表达平滑肌特征性蛋白SMA、sMHC和Calponin的表达。经VEGF165诱导，小鼠心脏Sca-1＋细胞同样发生了形态改变，上调内皮细胞特征基因CD31、vWF和VE-Cadherin的mRNA表达，同时表达内皮细胞特征性抗原CD31。结论：成体小鼠心脏Sca-1＋细胞在体外多种因素的分别作用下，能向心肌样细胞、平滑肌样细胞和内皮样细胞分化，具有心脏干细胞特性的多向分化潜能。%Objective:To study the differentiation potential of Sca-1+cells from adult mouse heart in vitro.Methods:We isolate Sca-1+ cells derived from mouse heart by means of immunomagnetic cell sorting.By using three different kinds of differentiation inducing factors,including BMP-2/FGF-4,TGF-β1 and VEGF165 ,to induce Sca-1+ cells to differentiated into the heart"three lines"invitro.The differentiation potential of the cells was identified from the changes of cell morphology, RT-PCR and immunofluorescence staining.Results:After BMP-2/FGF-4 induction,the mouse cardiac Sca-1+cells underwent morphological changes,up regulation of the mRNA expression ofα-MHC,β-MHC,MLC-2a and MLC-2v,and the protein expression of cTNT and cMHC,which were the specific markers of cardiomyocytes.After TGF-β1 induction,these Sca-1+cells also had morphological changes,up regulation of mRNA expression ofα-SMA and Calponin,and the protein expression of SMA,sMHC,and Calponin,which were the specific markers of smooth muscle cells.After VEGF165 induction,the Sca-1+cells also underwent morphological changes,up regulation of endothelial cell specific gene CD31,vWF,and VE-Cadherin expression,and the expression of endothelial cell specific antigen CD3 1 .Conclusions:Adult mouse cardiac Sca-1+ cells can differentiate into cardiomyocyte-like cells,smooth muscle-like cells and endothelial-like cells under different inducing factors in vitro,which suggest these cells have multi-lineage differentiation potential as cardiac stem cells.
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