绿色荧光蛋白转基因小鼠脂肪间充质干细胞的多向分化潜能  

Multi-lineage potential of adipose tissue-derived stromal cells from green fluorescent protein transgenic mice

作　　者：孙雅娟[1] 刘鲁川[1] 金岩[2] 邓蔓菁[1] 方军[2] 陆伟[2] 杨振华[2] 

机构地区：[1]第三军医大学大坪医院野战外科研究所口腔科,重庆400042 [2]第四军医大学组织工程研发中心,陕西西安710033

出　　处：《第四军医大学学报》2008年第7期623-625,共3页Journal of the Fourth Military Medical University

基　　金：重庆市自然科学基金(CSTC;2007BB5057)

摘　　要：目的:研究绿色荧光蛋白(GFP)转基因小鼠脂肪间充质干细胞(ADSC)体外定向诱导下的多向分化潜能.方法:取出生后4 d的GFP转基因小鼠腹股沟的皮下脂肪组织,经过胶原酶消化,加入脂肪组织来源干细胞条件培养基中培养.将生长状态良好的第5代GFP-ADSCs分别加入成骨及成脂条件培养液进行体外多向分化诱导研究.并分别采用茜素红钙盐染色和油红O染色进行鉴定.结果:成功获得了稳定表达GFP的ADSCs,GFP-ADSC经成骨诱导20 d后茜素红染色可见有橘红色钙盐沉积;经成脂诱导10 d后油红O染色阳性.结论:GFP作为报告基因在ADSCs分化过程中没有被关闭,亦未影响到ASCs的多向分化潜能,是ADSC在体多向分化研究的一个有效示踪工具.AIM： To study the multi-lineage potential of adipose tissue-derived stromal cells （ADSCs） from transgenic mice with green fluorescent protein （GFP） gene. METHODS： Adipose tissues were isolated from the groin region of a 4-day-old GFP transgenic mouse, and incubated in a collagenase solution, then cultivated in conditioned culture medium to obtain GFP-ADSCs. The passage 5 GFP-ADSCs were induced to differentiate into osteoblasts, or adipocytes with solution of calcium induction medium or adipogenic medium respectively. After calcium induction, the alizarin red staining was performed to test the formation of calcium concentration. The adipo-induced MSCs were detected with oil red 0 staining. RESULTS： ADSCs, stably expressing GFP, were obtained successfully from the transgenic mice with GFP gene. After 20 d calcium induction, jacinth calcium salts could be detected in the cells by alizarin red staining. At day 10 after being adipo-induced, the cells showed positive oil red 0 staining. CONCLUSION： The GFP, as report gene, is not closed during the differentiation, and exerts no effect on the multi-lineage potential of ADSCs, which can be used as an efficient tracking facility for studying the mechanism of multi-lineage potential of the ADSCs in vivo.

关 键 词：绿色荧光蛋白 转基因小鼠 脂肪间充质干细胞 多向分化潜能 

分 类 号：R318.5[医药卫生—生物医学工程]