小鼠心源性CD51阳性细胞的分离与鉴定  

Isolation and Determination of Characteristics of CD51^+ Cells in Myocardial Tissue of Mouse

作　　者：陈阳[1] 李桂兰[2] 姜美花[3] 彭朝权[1] 

机构地区：[1]中山大学附属第三医院心血管内科,广东广州510630 [2]中山大学中山医学院,广东广州510080 [3]中山大学中山医学院人体解剖学教研室,广东广州510080

出　　处：《中山大学学报（医学科学版）》2014年第3期424-430,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81370214);广东省自然科学基金(S2013010016392)

摘　　要：【目的】探讨整合素蛋白αv(CD51)阳性细胞在小鼠正常心脏组织中的分布情况并鉴定其是否存在心肌干细胞相关特性。【方法】通过免疫荧光染色,RT-PCR,Q-PCR等方法,观察CD51+细胞在C57小鼠心脏中的表达及分布情况;应用流式细胞仪进一步检测心脏中CD51的表达比例,并分选出CD51阳性细胞进行体外培养;利用CCK-8实验检测细胞增殖能力;细胞稳定传代后进行成骨、成脂诱导,检测其多向分化能力。【结果】实验结果显示小鼠心脏中存在CD51+细胞,随年龄增大其表达水平呈下降趋势;在出生7d C57小鼠心脏组织中可观察到CD51+细胞广泛分布于心房、心室;流式细胞仪分选出生7 d C57小鼠心脏组织中阳性细胞结果显示阳性率约4%左右;诱导结果显示该细胞可以分化形成骨及脂肪组织,进一步证明其存在多向分化能力。【结论】小鼠心肌组织中CD51+细胞是具有干细胞性的细胞亚群,可能为心脏来源的MSC样干细胞。[Objective] To isolate and culture intergrin αv （CD51＋） positive cells in mouse heart,and verify its characteristics of cardiac stem cell for investigation of its distribution.[Methods] To characterize CD51 ＋ cell expression and distribution in postnatal C57 mouse heart by using RT-PCR,Q-PCR,and confocal microscopy analysis.The CD51＋ cell proliferation and multi-differentiation capacity were tested by CCK-8 analysis and osteogenic,adipogenic induction analysis,respectively.[Results] Experimental results confirmed that the CD51 ＋ cells were widely expressed in the atrium and ventricle,and which expression level declined with the age.Positive cells were sorted by flow cytometry and showed that the positive rate was approximately 4％ in 7 d C57 mouse.Cultured CD51＋ cells showed a strong ability of proliferation and adipogenic/osteogenic differentiation.[Conclusion] CD51 ＋ cells existed in myocardial tissue of mice are a cell subsets with stem cell characteristics,it may be a category of heart derived MSC-like stem cells.

关 键 词：心肌干细胞 CD51 流式分选 增殖 多向分化 

分 类 号：Q254[生物学—细胞生物学]