目的：分离、扩增干细胞抗原1（ Sca-1）阳性心脏内源性干细胞，并观察其标记物情况。方法取C57 BL／6小鼠心肌组织，用磁珠分选法分离Sca-1阳性内源性心脏干细胞，并对其传代扩增。观察初分离及传代过程中细胞形态的变化，用流式细胞仪检测初分离及扩增后（第5代） Sca-1阳性心脏干细胞阳性率，以RT-PCR法检测Sca-1阳性及阴性心脏干细胞肌球蛋白重链6（MYH6）、肌钙蛋白T（cTnT）、心肌转录因子4（GATA4）标记物的表达。结果初分离的Sca-1阳性心脏干细胞为圆形，比血细胞小；传代后细胞呈现星形，伸出较长的伪足，第5代后形状差异不大。原代、第5代心肌细胞Sca-1阳性心脏干细胞阳性率分别为81．0％±10．4％、90．0％±5．0％，P＞0．05；Sca-1阳性心脏干细胞MYH6、cTnT、GATA4相对表达量分别为0．50±0．21、0．72±0．04、0．72±0．05，阴性心脏干细胞分别为0．20±0．02、0．86±0．06、0．71±0．05，两者MYH6、cTnT比较，P均＜0．05。结论成功分离出Sca-1阳性心脏干细胞，且传代稳定、纯度较高，高表达MYH6。%Objective To isolate and amplify the Sca-1 positive cardiac stem cells from C57BL/6 mice in vitro and to observe its markers .Methods The myocardial tissues from C 57 BL/6 mice were selected , and we separated the Sca-1 pos-itive cells by the magnetic cell sorting and amplified its passage .We observed the morphology changes in the culture process.The flow cytometry was used to test the Sca-1 +purity of the primary and fifth generation of myocardial cells .RT-PCR was used to detect the expression of multiple factors (MYH6, cTnT and GATA4) in the isolated Sca-1 positive cells and negative cells .Results The primary cultured Sca-1 positive cells were round , smaller than blood cells;the sub-cul-tured were starlike , and the pseudopodia were long , and after the fifth generation , the morphology of cells showed little difference.The Sca-1 +positive rates of primary and the fifth generation were 81%±10.4% and 90%±5%, separately (P>0.05).The relative expression of MYH6, cTnT and GATA4 in Sca-1 +cells was 0.5 ±0.21, 0.72 ±0.04 and 0.72 ±0.05, separately;while the expression in the Sca-1 -cells was 0.2 ±0.02, 0.86 ±0.06 and 0.71 ±0.05.Significant difference was found in the expression of MYH6 and cTnT (all P<0.05).Conclusion Sca-1 positive cardiac stem cells can be isolated successfully and sub-cultured stablely.Furthermore, Sca-1 +cells show high expression of MYH6.
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