猕猴脂肪干细胞体外多向分化潜能的实验研究  

In vitro multipotent differentiation capability of adipose-derived stromal cells from a rhesus monkey

作　　者：黄喜军[1] 朱庆棠[1] 江丽 何彩凤[1] 郑灿镔[1] 刘小林[1] 

机构地区：[1]中山大学附属第一医院显微创伤骨科,广州510080 [2]骨科研究所

出　　处：《中华创伤骨科杂志》2014年第1期52-57,共6页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金（30700847,31070869）;广东省自然科学基金（9251008901000017）;广东省科技计划项目（20108031100006）

摘　　要：目的探讨猕猴来源的脂肪干细胞（ADSCs）的分离和培养方法及在体外多向分化的能力。方法健康成年雄性猕猴1只，年龄4岁，体质量4．5妇。从该猕猴胭窝深部脂肪组织中分离出成纤维样细胞，观察其生长特性，采用流式细胞仪检测其细胞表面标志，利用相应的诱导液分别诱导其向成骨细胞、脂肪细胞、施万细胞方向分化；观察诱导后细胞的形态变化，并分别用茜素红染色、油红0染色及细胞免疫荧光法检测诱导效果。结果猕猴脂肪组织分离、培养出的成纤维样细胞增殖迅速；细胞表面标志CD34、CD14、CD71、CD106、CD117、CD31、CD45、CD80、CD86及人白细胞DR抗原呈阴性表达；CD29、CD105、CD73、CD90呈阳性表达。在相应的诱导体系下可向成骨细胞、脂肪细胞、施万细胞分化，其中成脂诱导率为47．9％～61．5％（平均54．7％）；成施万细胞诱导后ADSCs胞体呈细小梭形，伸出2—3条细长突起；高表达神经胶质细胞标志蛋白S-100和胶质纤维性酸性蛋白质，平均诱导率分别为66．5％和60．6％，双重表达率平均为41．2％。结论从猕猴脂肪组织中可以分离并培养出具有较强的自我更新及多向分化潜能的ADSCs，在体外诱导条件下能分化为类施万细胞。Objective To study the in vitro multipotent differentiation capacity of adipose-derived stem ceils （ADSCs） which were isolated and cultivated from a rhesus monkey. Methods Used for this study was a healthy adult male rhesus monkey, 4 years of age and 4.5 kg in weight. After spindle-like ADSCs were isolated from the popliteal fossa fat of the monkey, they were cultured and passaged for observation of their biological characteristics. The expression of cell surface markers was detected by flow cytometer. After the ADSCs were induced to differentiate into osteoblasts, adipocytes and Schwann cells by corresponding in- duction medium respectively, their morphological features after induction were observed by alizarin red staining, oil red O staining and immunofluorescence assay respectively. Results The ADSCs isolated and cultured from the rhesus monkey had a strong ability of proliferation in a spindle-like shape, negative for CD34, CD14, CD71, CDI06, CDll7, CD31, CD45, CD80, CD86 and human leukocyte antigen-DR （HLA-DR） HLA-DR but positive for CD29, CD105, CD73 and CD90. They could differentiate respectively into osteoblasts, adipocytes and Schwann cells in the corresponding induction system. Their adipogenic rate was from 47.9% to 61.5% （average, 54. 7% ） . After induced into Schwann cell-like cells （SCLCs）, the ADSCs were similar to Schwann cells in that they had small and spindle nucleus and 2 to 3 elongated axons and high expression of S-100 and glial fibrillary acidic protein（GFAP）. The average positive percentages of S-100 and GFAP expressions were 66.5% and 60. 6% , respectively, and the double positive percentage was 41.2%. Conclusion The ADSCs derived from a rhesus monkey have a strong capacity of self-renewal and multipotent differentiation so that they can differentiate into SCLCs in vitro.

关 键 词：干细胞 细胞分化 组织工程 猕猴属 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]