骨髓间充质干细胞异体移植对小鼠实验性自身免疫性脑脊髓炎的治疗作用
发布时间：2019-08-01 11:25
【摘要】：目的探讨大鼠骨髓间充质干细胞(BMSCs)异体移植对小鼠实验性自身免疫性脑脊髓炎(EAE)的治疗作用。方法全骨髓贴壁培养法获得大鼠BMSCs,流式细胞术检测细胞免疫表型,并诱导成骨方向分化。取雌性C57BL/6小鼠,随机分为3组:正常对照组、PBS组和大鼠BMSCs组,用髓鞘少突胶质细胞糖蛋白(MOG)35~55联合完全弗氏佐剂诱导建立EAE模型。小鼠免疫后38d和48d腹腔注射PBS或大鼠BMSCs进行治疗,神经功能评分观察各组小鼠神经功能变化。二次治疗12d后取各组小鼠脊髓、脾脏和外周血。HE染色及Luxol fast blue染色观察脊髓炎性细胞浸润及髓鞘脱失情况;脾脏制成单细胞悬液,细胞CFSE标记后10mg/L刀豆球蛋白(Con A)和MOG_(35~55)刺激培养3d,观察脾细胞增殖情况。ELISA检测大鼠BMSCs移植后外周血细胞因子干扰素γ(IFN-γ)、白细胞介素17(IL-17)含量。结果流式细胞术显示,大鼠BMSCs第3代(P3)细胞表达抗原CD29、CD90、CD106,不表达CD45。体外诱导其能向成骨分化。小鼠发病后,大鼠BMSCs组小鼠神经功能缺损症状较PBS组减轻,评分降低。HE染色和Luxol fast blue染色结果显示,大鼠BMSCs组脊髓炎细胞浸润和脱髓鞘比同时间点PBS组减轻(P0.05)。Con A和MOG_(35~55)刺激培养后,PBS组和大鼠BMSCs组脾细胞增殖增加,而大鼠BMSCs组又较PBS组降低。与PBS组相比,大鼠BMSCs组血浆细胞因子IFN-γ、IL-17含量降低(P0.05)。结论全骨髓贴壁培养法能有效分离纯化BMSCs,大鼠BMSCs异体移植对小鼠EAE模型有治疗作用。
[Abstract]:Objective To study the effect of bone marrow mesenchymal stem cells (BMSCs) on experimental autoimmune encephalomyelitis (EAE) in mice. Methods BMSCs and flow cytometry were used to detect the cell-immune phenotype and to induce the differentiation of the osteogenesis. The female C57BL/6 mice were randomly divided into three groups: the normal control group, the PBS group and the rat BMSCs group, and the EAE model was established by the combination of the myelin oligodendrocyte glycoprotein (MOG)35-55 and the complete Freund's adjuvant. After the mice were immunized for 38d and 48d, the rats were injected with PBS or rat BMSCs for treatment, and the neurological function scores of the mice were observed to observe the changes of the neurological function in each group. The spinal cord, spleen and peripheral blood of each group were taken after 12 days of secondary treatment. The proliferation of spleen cells was observed by HE staining and Luxol fast blue staining to observe the infiltration of myelitis and the depigmentation of the pulp. The spleen was made into single cell suspension, and the cell CFSE was labeled with 10 mg/ L of concanavalin (Con A) and MOG _ (35-55). The levels of IL-17 and IL-17 in peripheral blood of rat BMSCs were detected by ELISA. Results Flow cytometry showed that the expression of CD29, CD90 and CD106 in the third generation (P3) cells of rat BMSCs did not express CD45. In vitro induce it to differentiate into osteogenesis. After the onset of the mouse, the neurological deficit in the BMSCs group was reduced in the group of PBS and the score was decreased. The results of HE staining and Luxol fast blue staining showed that the proliferation and proliferation of the spleen cells in the PBS group and the rat BMSCs group increased after the rats BMSCs group and the rats BMSCs group were stimulated and cultured, and the BMSCs group in the rat was lower than the PBS group. Compared with the PBS group, the content of IFN-1 and IL-17 in the rat BMSCs group decreased (P0.05). Conclusion The whole bone marrow adherent culture method can effectively separate and purify BMSCs, and the rat BMSCs allografts have a therapeutic effect on the mouse EAE model.
【作者单位】： 贵州医科大学基础医学院组织学与胚胎学教研室;贵州医科大学贵州省细胞工程生物医药技术国家地方联合工程实验室贵州省再生医学重点实验室;
【基金】：贵州省教育厅基金项目[黔教合KY字2016(071)] 贵州省科技厅优秀青年科技人才培养对象专项基金[黔科合人字(2015)07号] 贵州省科技合作项目联合基金[黔科合LH字(2016)7363]
【分类号】：R457.7;R744.5
本文编号：2521720
[Abstract]:Objective To study the effect of bone marrow mesenchymal stem cells (BMSCs) on experimental autoimmune encephalomyelitis (EAE) in mice. Methods BMSCs and flow cytometry were used to detect the cell-immune phenotype and to induce the differentiation of the osteogenesis. The female C57BL/6 mice were randomly divided into three groups: the normal control group, the PBS group and the rat BMSCs group, and the EAE model was established by the combination of the myelin oligodendrocyte glycoprotein (MOG)35-55 and the complete Freund's adjuvant. After the mice were immunized for 38d and 48d, the rats were injected with PBS or rat BMSCs for treatment, and the neurological function scores of the mice were observed to observe the changes of the neurological function in each group. The spinal cord, spleen and peripheral blood of each group were taken after 12 days of secondary treatment. The proliferation of spleen cells was observed by HE staining and Luxol fast blue staining to observe the infiltration of myelitis and the depigmentation of the pulp. The spleen was made into single cell suspension, and the cell CFSE was labeled with 10 mg/ L of concanavalin (Con A) and MOG _ (35-55). The levels of IL-17 and IL-17 in peripheral blood of rat BMSCs were detected by ELISA. Results Flow cytometry showed that the expression of CD29, CD90 and CD106 in the third generation (P3) cells of rat BMSCs did not express CD45. In vitro induce it to differentiate into osteogenesis. After the onset of the mouse, the neurological deficit in the BMSCs group was reduced in the group of PBS and the score was decreased. The results of HE staining and Luxol fast blue staining showed that the proliferation and proliferation of the spleen cells in the PBS group and the rat BMSCs group increased after the rats BMSCs group and the rats BMSCs group were stimulated and cultured, and the BMSCs group in the rat was lower than the PBS group. Compared with the PBS group, the content of IFN-1 and IL-17 in the rat BMSCs group decreased (P0.05). Conclusion The whole bone marrow adherent culture method can effectively separate and purify BMSCs, and the rat BMSCs allografts have a therapeutic effect on the mouse EAE model.
【作者单位】： 贵州医科大学基础医学院组织学与胚胎学教研室;贵州医科大学贵州省细胞工程生物医药技术国家地方联合工程实验室贵州省再生医学重点实验室;
【基金】：贵州省教育厅基金项目[黔教合KY字2016(071)] 贵州省科技厅优秀青年科技人才培养对象专项基金[黔科合人字(2015)07号] 贵州省科技合作项目联合基金[黔科合LH字(2016)7363]
【分类号】：R457.7;R744.5
【相似文献】
相关期刊论文 前5条
1 源淑英;陈国锐;;带血管甲状旁腺异体移植手术前后的护理[J];中华护理杂志;1988年04期
2 牟善初,王新明,敖建华,周桂芳;经静脉超声造影二次谐波成像对异体移植肾脏的研究[J];中国超声医学杂志;2000年11期
3 谢丽萍,张悦;异体角膜缘干细胞移植术的护理[J];中华护理杂志;2001年03期
4 谭琳玲,张军花,廖晓艳,康卫平;3例异体手移植术的护理配合[J];南方护理学报;2004年07期
5 ;[J];;年期
相关重要报纸文章 前2条
1 朴燕;双臂异体移植首获成功[N];医药经济报;2002年
2 ;世界首例双手臂异体移植手术成功[N];光明日报;2002年
，本文编号：2521720
本文链接：https://www.wllwen.com/yixuelunwen/shenjingyixue/2521720.html
最近更新
教材专著
