论文部分内容阅读
目的:了解大鼠外胚间充质干细胞(EMSC)的生物学特性,探索体内输入EMSC能否促进照射动物造血功能的恢复。方法:取E11.5SD大鼠胚胎颌突,消化法培养获得外胚间充质干细胞,倒置显微镜下观察细胞形态和生长状况,免疫组化鉴定细胞来源;雄性SD大鼠(180～220g)分为外胚间充质干细胞组(A组)、照射对照组(B组)、成纤维细胞组(C组)、正常对照组(D组),前三组给予60Coγ射线全身照射,剂量6.0cGy,动态观察大鼠外周血变化,并于第4周检测骨髓有核细胞数量;测定大鼠骨髓细胞粒—巨噬细胞(CFU-GM)形成能力;采用内源性脾结节法测定脾集落形成能力。结果:培养的细胞呈单层生长,成纤维状、长梭形、有2～4个突起,抗HNK1、Vi mentin、S-100、NSE染色阳性,抗GFAP、NF、MA染色阴性,证实为未分化外胚间充质细胞;体内植入EMSC能够明显促进大鼠放射后外周血白细胞和血红蛋白的恢复,增加骨髓有核细胞数,减少骨髓造血细胞的坏死、凋亡,促进骨髓细胞的CFU GM的形成率并能有效地支持造血细胞增殖、分化。结论:EMSC具有干细胞特征,能促进照射大鼠造血功能的恢复。
OBJECTIVE: To understand the biological characteristics of rat ectomesenchymal stem cells (EMSCs) and to explore whether in vivo EMSCs can promote the recovery of hematopoietic function in irradiated animals. Methods: Eighteen embryos of E11.5SD rat embryos were implanted into the ectodermal stem cells by digestion method. Morphology and growth of the cells were observed under an inverted microscope. The cells were identified by immunohistochemistry. Male Sprague-Dawley rats (180-220 g) (Group A), irradiation control group (group B), fibroblasts group (group C) and normal control group (group D). The first three groups were given 60Co γ ray with body weight of 6.0 cGy , The changes of peripheral blood in rats were observed dynamically and the number of nucleated cells in bone marrow was measured at the fourth week. The ability of CFU-GM formation in rat bone marrow cells was measured. Spleen colonies were detected by endogenous spleen nodules Ability to form. Results: The cultured cells showed monolayer growth, fibroblast shape and long fusiform shape, with 2-4 protuberances, positive staining for anti-HNK1, Vimentin, S-100 and NSE, negative staining for anti-GFAP, NF and MA, Undifferentiated ectomesenchymal cells; EMSC implantation can significantly promote the recovery of peripheral blood leukocytes and hemoglobin after radiation in rats, increase the number of bone marrow nucleated cells, reduce the necrosis and apoptosis of bone marrow hematopoietic cells, and promote the CFU of bone marrow cells GM formation rate and can effectively support hematopoietic cell proliferation and differentiation. Conclusion: EMSC has stem cell characteristics and can promote the recovery of hematopoietic function in irradiated rats.