维甲酸对脂肪源干细胞中生殖标记表达的影响  

Effect of retinoic acid on alkaline phosphatase expression in rat adipose-derived stem cells

作　　者：段富华[1] 杨春[1] 杨会营[1] 余美春[1] 陶晖[1] 曾文钦[1] 戴景兴[1] 原林[1] 

机构地区：[1]南方医科大学人体解剖学教研室,广东广州510515

出　　处：《南方医科大学学报》2012年第1期32-36,共5页Journal of Southern Medical University

基　　金：国家重点基础研究973计划(2007CB512705);国家自然科学基金(30801464);广东省高新技术产业化项目-工业攻关"自体筋膜组织移植系统的研发(2011B010500004)~~

摘　　要：目的探索脂肪源干细胞(ADSCs)在维甲酸(RA)诱导下是否表达原始生殖细胞标记碱性磷酸酶(ALP)及生殖标记基因vasa。方法分离培养2个月龄SD雌性大鼠ADSCs,传至第3代进行成脂、成骨诱导和表面标记鉴定。将第3代ADSCs分别用10-5、10-6、10-7mol/L RA诱导7 d和14 d,采用ALP检测试剂盒检测各组ALP的活性水平。用RT-PCR方法检测10-5mol/L RA组Vasa mRNA表达。结果将第3代ADSCs分别用10-5、10-6、10-7mol/L RA诱导7 d的D520 nm值分别为0.59±0.04,0.27±0.07,0.15±0.03,对照组为0.07±0.01,诱导14 d的OD值分别为0.42±0.02,0.34±0.01,0.19±0.02,对照组为0.07±0.01,说明RA能显著性促进ADSCs碱性磷酸酶的表达(P<0.01)。第3代ADSCs用10-5mol/L RA诱导7 d Vasa mRNA的表达为阴性。结论第3代ADSCs在维甲酸诱导下能够表达原始生殖细胞标记碱性磷酸酶,但用10-5mol/L RA诱导7 d不能表达生殖标记基因Vasa。Objective To investigate whether adipose-derived stem cells（ADSCs） induced by retinoic acid（RA） in vitro express primordial germ cell marker alkaline phosphatase（ALP） and vasa.Methods ADSCs were isolated from adult female SD rats and cultured in vitro.The third passage of ADSCs was identified by adipogenic differentiation,osteogenic differentiation and cell surface marker detection.The ADSCs were treated with 1×10-5,1×10-6,or 1×10-7 mol/L RA for 7 or 14 days,and the cellular expression of ALP was detected.vasa mRNA expression in ADSCs treated with 1×10-5 mol/L RA for 7 days was detected using RT-PCR.Results The OD value of ADSCs treated with 1×10-5,1×10-6,or 1×10-7 mol/L RA was 0.59±0.04,0.27±0.07,and 0.15±0.03 after a 7-day treatment,and was 0.42±0.02,0.34±0.01,and 0.19±0.02 after a 14-day treatment,respectively,demonstrating significantly enhanced ALP expression in RA-treated ADSCs compared with that in the control cells（0.07±0.01 and 0.07±0.01 at 7 and 14 days,respectively,P0.01）.The ADSCs showed a negative vasa mRNA expression after 1×10-5 mol/L RA treatment for 7 days.Conclusion RA-induced ADSCs express ALP,a marker of primordial germ cells,but does not express the primordial germ cell marker vasa.

关 键 词：脂肪源干细胞 维甲酸 原始生殖细胞 碱性磷酸酶 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]