摘要
目的 初步验证量子点能否对共培养睾丸体细胞进行标记,为睾丸体细胞的体内外研究提供一种细胞损伤小,且简便、有效的标记手段.方法 取雄性Wistar大鼠的睾丸组织小块,胶原酶消化,差速贴壁法收集睾丸体细胞,取第一代睾丸体细胞以0.25%胰酶蛋白酶-0.02%EDTA消化,制备单细胞悬液,与量子点共同孵育2h后贴壁及传代培养,共聚焦荧光显微镜动态观察细胞生长、增殖情况.结果 相差显微镜下观察,标记细胞形态无明显改变,荧光显微镜下可见摄取量子散在分布于细胞内,主要集中于细胞核周围,分裂细胞及增殖活跃细胞摄取量明显高于老化细胞.传代后仍可见细胞内的量子点分布,荧光强度未见明显衰减.结论 量子点能够被共培养睾丸体细胞摄取,且摄取速度快,荧光显微镜下易于观察.量子点标记法对共培养细胞损伤小,操作简便易行,是共培养睾丸体细胞标记的理想选择之一.
Objective To study the Quantum dots （QDs） to label testis somatic cells, try to find a more capable and establish basis for the labeled cells in the transplantation study of the co-cultured testis somatic cells. Methods A piece of tissue of testes of male wistar rat was got, digested by eollagenase, isolation and purification by different attachment method. The cells got by this method were co-cuhured testis somatic cells. The first generation cells were digested by 0.25% trypsinase-0.2% EDTA, then mixed with the QDs and cultured for 2 hours. The labeled cells were cultured and went down to the future generation. Observation was performed under microscope. Results By microscope, we observed that the cells grew well. The QDs entered the cells and scatted in the cells. As time by, the QDs were gathered around the cell core. The younger cells got more QDs than the older ones. The QDs in the cells were inherited by the daughter cells： The quantum dots in the cells were still brighter. Conclusion The co-cultured testis somatic cells could be labeled quickly with the QDs, and easy to be observed under microscope. It is an easier and less damage method to label the cells than before. And the QDs labeling is one of the best choose for monitoring of the co-cultured testis somatic cells.
出处
《中国美容整形外科杂志》
CAS
2011年第3期164-167,共4页
Chinese Journal of Aesthetic and Plastic Surgery
关键词
量子点
共培养
睾丸体细胞
细胞标记
Quantum dots
Co-cultured
Testis somatic cells
Cell labeling
