解剖学报 ›› 2013, Vol. 44 ›› Issue (3 ): 339-344.doi: 10.3969/j.issn.0529-1356.2013.03.009

• 细胞和分子生物学 • 上一篇    下一篇

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

齐立杰郭康 李琼1,3  郭志坤1*    

  1. 1. 新乡医学院河南省医用组织再生重点实验室 ; 2. 第三附属医院肿瘤科,河南 新乡 453003; 3.华中科技大学同济医学院组织学与胚胎学教研室,武汉 430000
  • 收稿日期:2012-08-17 修回日期:2012-11-23 出版日期:2013-06-06 发布日期:2013-07-16
  • 通讯作者: 郭志坤 E-mail:gzk@xxmu.edu.cn
  • 基金资助:

    省自然科学基金资助项目

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

QI Li-jieGUO Kang2  LI Qiong 1,3  GUO Zhi-kun 1*   

  1. 1.Key Open Laboratory for Medical Tissue Regeneration of He’nan Province,Xinxiang Medical University, He’nan Xinxiang 453003, China;
    2. Department of Oncology,the Third Affiliated Hospital,He’nan Xinxiang 453003, China; 3. Department of Histology and Embryology,
    Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430000, China
  • Received:2012-08-17 Revised:2012-11-23 Online:2013-06-06 Published:2013-07-16

摘要:

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。

关键词: CD73, 5-氮杂胞苷, 心肌组织裂解液, 脂肪源间充质干细胞, 细胞培养, 免疫荧光, 小鼠

Abstract:

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.

Key words: CD73, 5-aza, Myocardial tissue lysate, Adipose-derived mesenchymal stem cell, Cell culture, Immunofluorescence, Mouse