绿色荧光标记共培养睾丸体细胞体内-外培养观察.docVIP

绿色荧光标记共培养睾丸体细胞体内\外培养观察[摘要]目的：对Wistar大鼠睾丸间质体细胞转染携带绿色荧光蛋白基因的慢病毒，为进一步标记睾丸间质体细胞移植组织工程实验奠定基础。方法：获取Wistar大鼠睾丸间质体细胞，用慢病毒转载绿色荧光对其转染标记，体外培养，在共聚焦荧光显微镜下动态观察细胞生长、增殖等情况。回植于去势鼠体内，培养一定时期后取心脏血进行雄激素的检测。结果：睾丸间质体细胞于90h后，在荧光显微镜蓝光激发波长下，可发出明亮的绿色荧光，转染率达80%。传代后的细胞仍能发出明亮的绿色荧光。回植后血清学检测有明显高于对照组的激素检出。结论：经GFP转染后的睾丸间质体细胞仍具有睾丸间质体细胞的特性。采用携带绿色荧光蛋白基因的慢病毒标记睾丸体细胞中梭形及不规则形细胞（SLCs等）转染细胞效率高且持久、简便有效，且能保持睾丸间质体细胞原有生物学特性。 [关键词]绿色荧光；共培养睾丸体细胞；细胞标记 [中图分类号]Q813.1[文献标识码]A[文章编号]1008-6455（2011）04-0608-05 Detection of the culture in vitro of the co-cultured testis somatic cells and GFP labeling LI Ming1,XU Feng2,XU Hong-xia1,WANG Xiao-yun3,FAN Xing4,BI Hong-da3,XING Xin3 (1.Department of Plastic Surgery, Navy General Hospital,Beijing100037,China;2.Nuclear of Shanghai Ninth People’s Hospital Affiliated Shanghai Jiaotong University School of Medicine, Shanghai 200011,China;3.Department of Plastic Surgery,Changhai Hospital Affiliated The Second Military Medical University Shanghai 200433,China;4. The Institute of Plastic Surgery,PLA of The Fourth Military Medical University,Xi’an 710032,Shaanxi,China) Abstract:ObjectiveGreen fluorescent protein(GFP) are nanoparticles which have strong fluorescence intensity and more are resistant to photobleaching thanorganic fluorescence probes in cell biology and biomedicine. In this report, we labeled it with GFP for the co-cultured testis somatic cells.MethodsWe got seed-cells labeling with GFP. We cultured the labeled cells and went down to the future generation. We observed it by the micrscope. Assay the serum testeosterone level of the transplanted rats by heart puncture.ResultsThe co-cultured testis somatic cells labeled with GFP were easy to label. The GFP in the cells were brighter, more stable. The GFP in the cells were inherited by the daughter cells. Conclusion GFP labeling opened up new possibilities for monitoring of the co-cultured testis somatic cells. Especially for stem Leydig cells, SLC, Progenitor Leydig cells, PLC. And can remain its proper biolog