3种诱导法诱导CD73+脂肪间充质干细胞成心肌细胞效果的比较

目的 从脂肪来源的间充质干细胞（ADMSCs）的混合细胞群中分离纯化出CD73阳性的细胞亚群，证明几种不同诱导法诱导CD73细胞的
成心肌分化潜能。方法 体外分离培养1~3 月龄小鼠的ADMSCs，利用流式细胞仪从ADMSCs中分选出CD73⁺和CD73^-两个细胞亚群。分别培养两组细
胞，利用5-氮杂胞苷（5-aza）、心肌组织裂解液和5-aza+心肌组织裂解液对两组分选出的细胞分别进行成心肌的诱导。利用免疫细胞化学技术
检测3种诱导法的成心肌效果。结果 未分选的ADMSCs可分化为成脂和成骨细胞，分选的CD73⁺细胞具备分化为心肌细胞的良好潜能。3种诱导法
均能诱导CD73⁺ADMSCs向心肌方向分化，5-aza诱导CD73⁺ADMSCs分化为心肌细胞率为（22.99±6.72）%，心肌组织裂解液组心肌细胞率（14.12±5.42）%，5-aza+心肌组织裂解液组心肌细胞率（26.94±6.11）%。3种方法比较，5-aza+心肌组织裂解液的诱导效果最佳（P<0.05）。结论 CD73+比CD73-
的ADMSCs更易于分化为心肌细胞。化学诱导因素和体外模拟心肌微环境，可高效诱导脂肪间充质干细胞分化为心肌细胞。

Comparison of effects on the induction of three methods of inducing CD73 + adipose-derived mesenchymal stem cells into myocardial cells

Objective To isolate and purify CD73 positive cell subsets from the mixed adipose-derived mesenchymal stem cells
（ADMSCs）, and to compare the differentiation potential of CD73 cell to cardiomyocyte by three methods. Methods ADMSCs of
1-3-month-old mice were isolated and cultured in vitro. ADMSCs subpopulations for CD73⁺ and CD73^- were selected by flow cytometry,
and were cultured separately. Two groups of the sorted cells were induced into the myocardium by using 5-aza, myocardial tissue
lysate and 5-aza+ myocardial tissue lysate respectively. Immunohistochemistry was used to detect the effect of myocardium after
the induction of three ways. Results Unsorted ADMSCs differentiated into adipocytes and ostcoblasts. Sorted CD73+ADMSCs possessed
a perfect differentiating potential for cardiaomyocyte. All three induction methods inducted CD73+ ADMSC into cardiomyocytes.
The differentiation rate of myocardial cells which induced by 5-aza was (22.99±6.72)%, the rate by myocardium lysate was
(14.12±5.42)%, and the rate by 5-aza+myocardium lysate was (26.94±6.11)%. Of the three methods, 5-aza+myocardium lysate was
the best method for inducting CD73⁺ ADMSCs to myocardial cell(P<0.05). Conclusion CD73⁺ADMSCs may differentiate to cardiomyocyte
more easily than CD73^-ADMSCs. Chemical induction factors with the simulated myocardial microenvironment in vitro may efficiently
induce ADMSCs to differentiate into cardiomyocyte.