摘要
目的探讨骨髓间质干细胞的免疫原性,评估骨髓间质干细胞活化免疫系统的能力。方法分离培养大鼠股骨中的骨髓间质干细胞,扩增后分别经体外细胞刺激实验和体内移植后评估两个方面来评价大鼠骨髓间质干细胞的免疫原性。体外实验采用淋巴细胞激活实验进行,体内实验将骨髓间质干细胞经静脉移植到大鼠体内,观察移植后大鼠的脾脏指数,并解剖大鼠,了解骨髓间质干细胞的致瘤性。结果大鼠骨髓间质干细胞在体外增殖能力强,反复传代至10代以后,仍能良好生长,且生物学特征无明显变化。各组在加入CCK-8试剂终止反应后用酶标检测仪测得的结果显示:观察组为(1.515±0.737),阳性对照组为(1.966±0.421),阴性对照组为(1.302±0.259),三组比较差异具有统计学意义(P<0.05),观察组与阳性对照组比较差异具有统计学意义(P<0.05),阴性对照组与观察组比较差异无统计学意义(P>0.05)。细胞移植后,外源性骨髓间质干细胞进入大鼠血液系统,具有直接活化免疫系统的条件;同时随着循环系统迁移,向全身播散;在移植后不同时间点解剖大鼠,仔细观察各内脏器官、脑组织和骨骼肌,组织形态基本正常,未发现肿瘤形成。细胞移植后第8周处死大鼠,解剖分离脾脏,计算脾脏指数,结果显示,实验组大鼠脾脏指数(58.7±12.4)与对照组的(57.3±21.5)比较差异无统计学意义(P>0.05)。结论骨髓间质干细胞在体外共培养体系中不能活化淋巴细胞,经静脉移植后在体内也不能上调宿主的免疫状态,且无明显致瘤性,是用于细胞移植治疗的良好种子细胞。
Objective To investigate the immunogenicity of bone marrow mesenchymal stem cells and evaluate their ability to activate the immune system. Methods Bone marrow mesenchymal stem cells were isolated and cultured in vitro. After amplification, the immunogenicity of bone marrow mesenchymal stem cells was evaluated from both in vitro and in vivo. In vitro experiments were carried out using lymphocyte activation assay,and in vivo experiments were performed by transplanting bone marrow mesenchymal stem cells into rats. The spleen index of the transplanted rats was observed, and the rats were dissected to understand the tumorigenicity of bone marrow mesenchymal stem cells. Results The proliferation ability of bone marrow mesenchymal stem cells in rats is strong in vitro, and they can grow well after 10 generations, and there is no obvious change in biological characteristics. After the termination of reaction with CCK-8 reagent, the results measured by enzyme-linked detector in each group showed that: the observation group was(1.515±0.737), the positive control group was(1.966±0.421), the negative control group was(1.302±0.259), and the difference was statistically significant significant among the three groups(P<0.05). The observation group and the positive control group had statistically significant difference(P<0.05), and the negative control group and the observation group had statistically significant difference(P<0.05). After cell transplantation, the exogenous mesenchymal stem cells entered the blood system of rats, which had the conditions of activating the immune system directly;at the same time, they spread to the whole body with the circulation system migration;after transplantation, the rats were dissected at different time points, and the visceral organs, brain tissues and skeletal muscles were observed carefully. Their tissue morphology was basically normal, and no tumor was found. The rats were killed at 8 weeks after cell transplantation, the spleen was dissected and the spleen index was calculated. The results showed that the spleen index of the experimental group(58.7±12.4) was not significantly different from that of the control group(57.3±21.5)(P>0.05). Conclusion Bone marrow mesenchymal stem cells cannot effectively activate lymphocyte cell in vitro, and it does not significantly up-regulate the host’s immune status in vivo, and has no obvious tumorigenic performance. It is a good seed cell selection for cell transplantation therapy.
作者
梁爽
张贵育
孔杰
LIANG Shuang;ZHANG Gui-yu;KONG Jie(Shenzhen Baoan Hospital of Traditional Chinese Medicine,Shenzhen 518133,China)
出处
《中国实用医药》
2020年第2期195-198,共4页
China Practical Medicine
基金
广东省医学科研基金资助项目(项目编号：B2017031)
关键词
骨髓间质干细胞
免疫原性
细胞移植
Bone marrow mesenchymal stem cells
Immunogenicity
Cell transplantation
