经诱导的绿色荧光蛋白转基因C57BL雄鼠脾细胞移植重建雌鼠造血功能研究  

Transplantation of fish oocytes extracts-induced spleen cells from green fluorescent protein transgenic male C57BL mice to female mice for hematopoietic reconstruction

作　　者：阮光萍[1] 姚翔[1] 王金祥[1] 邓永丽[1] 庞荣清[1] 王强[1] 潘兴华[1] 

机构地区：[1]成都军区昆明总医院干细胞与组织器官工程研究中心,昆明650032

出　　处：《中华移植杂志（电子版）》2011年第4期28-31,共4页Chinese Journal of Transplantation(Electronic Edition)

基　　金：国家自然科学基金(31172170);中国博士后科学基金特别资助(201104748)

摘　　要：目的探讨雄性C57BL绿色荧光蛋白(GFP)鼠诱导的脾细胞移植对造血衰竭小鼠造血重建的作用。方法建立小鼠造血衰竭模型,移植鱼卵提取物诱导的雄性C57BL荧光鼠脾细胞,检测诱导后细胞的干细胞标志抗原表达;观察受体存活时间,检测外周血白细胞计数、外周血GFP阳性细胞,进行荧光原位杂交检测Y染色体;各组受体脾和肺切片观察GFP细胞分布。结果鱼卵提取物诱导的雄性C57BL荧光鼠脾细胞比未诱导的脾细胞表达更多的干细胞标志抗原。1×106个脾细胞经尾静脉回输经致死剂量照射的雌性小鼠,明显延长小鼠存活时间,提高小鼠外周血白细胞计数。诱导组受体外周血中检测到GFP阳性细胞,骨髓中检测出60%的细胞含Y染色体。诱导组受体脾和肺切片观察到GFP阳性细胞分布。结论鱼卵提取物诱导的小鼠脾细胞中含较多的多能干细胞,回输给造血衰竭小鼠能重建其造血功能。Objective To investigate the role of fish oocytes extracts induced spleen cells from green fluorescent protein （CFP） male C57BL mice in hematopoietic reconstitution of hematopoietic failure female mice. Methods The model of hematopoietic failure mice was established. The spleen cells of male fluorescent C57BL mice induced by fish oocytes extracts were transplanted to female mice. The expression of stem cell marker antigens on the induced cells was detected. Survival time of recipient mice was observed, peripheral white blood cells were counted, GFP positive peripheral blood cells were detected, and Y chromosome was detected by fluorescence in situ hybridization （FISH）. The spleen and lung biopsies of the recipient mice were done for observation of fluorescent cell distribution. Results The spleen cells of male C57BL mice induced by fish oocytes extracts expressed more stem cell marker antigens than the spleen cells without induction. A dose of 1 x 106 spleen cells infused via the tail vein back to the lethally irradiated female mice significantly prolonged survival time and increased the number of peripheral blood white blood cells. GFP positive cells were detected in recipient mice. In female recipient mice, FISH detected 60% of bone marrow cells with Y chromosomes. In the spleen and lung biopsy slices of mice in the induced group, distributions of fluorescent cells were observed. Conclusion The spleen cells induced by fish oocytes extracts contained more pluripotent stem cells than those without induction. The induced spleen cells infused to hematopoiefic failure mice can reconstruct the hematopoiefic function in mice.

关 键 词：转基因鼠 绿色荧光蛋白 诱导性多能干细胞 移植 造血重建 荧光原位杂交 

分 类 号：R392.1[医药卫生—免疫学]