异基因骨髓间充质干细胞移植在胶原诱导性关节炎大鼠体内的归巢

目的 通过对类风湿关节炎(RA)动物模型胶原诱导性关节炎(CIA)大鼠的实验研究,观察异基因骨髓间充质干细胞(MSCs)移植在CIA大鼠免疫器官和关节炎症部位的归巢和分布,并探讨其修复损伤关节的可能机制.方法 采用密度梯度离心结合贴壁培养法体外分离、培养大鼠MSCs,细胞表型鉴定;荧光染料PKH-26和5-溴2-脱氧尿嘧啶核苷(Brdu)标记MSCs,将64只大鼠随机分成正常组和CIA组,MSCs移植后第3、11、30、42天分别处死8只大鼠,取不同分组大鼠的胸腺、脾脏和关节组织制作病理切片,共聚焦显微镜联合免疫组织化学技术,观察MSCs移植后不同时点在不同脏器的迁移和分布,采用SPSS 12.0软件进行独立样本t检验.结果 异基因骨髓MSCs在CIA大鼠的脾脏、胸腺、关节炎症部位可较长期(42 d)的存在,MSCs移植后42 d,脾脏平均灰度值CIA组37.5±8.8高于正常组16.0±2.3,胸腺平均灰度值CIA组29.9±5.9高于正常组13.2±4.3,关节组织的平均灰度值CIA组78±8低于正常组93±14,差异均有统计学意义(P<0.05).结论 异基因骨髓MSCs移植优先定位于受损伤的组织和器官而发挥其治疗作用.

Abstract：

Objective To observe homing of mesenchymal stem cells (MSCs) in immune organs and inflammatory joints in collagen induced arthritis (CIA) rats. Methods Rats MSCs were isolated and expanded from bone marrow cells by density gradient centrifugation and adhering to the culture cell walls, and the phenotypes were assessed by flow cytometry. MSCs were labeled by PKH-26 and Brdu. Sixty-four rats were randomly divided into normal group and CIA group. Every 8 rats were sacrificed at 3, 11, 30, 42 days after transplantation of MSCs. At the end of the experiment, the specimens of thymus gland, spleen, ankle joints were exposed, fixed, decalcified, wrapped and cut into slices. Confocal laser scanning microscope and immunohistochemical method were used to observe migration and distribution of MSCs in different organs. Independent samples group t test with SPSS 12.0 software package was used for statistical analysis. Results It was found that allogenic MSCs could stay in spleen, thymus gland and joints of CIA rats for a relatively long period (42days). Forty-two days after transplantation of MSCs, the average grey scale values of spleen and thymus gland in CIA group (37.5±8.8, 29.9±5.9 respectively) were significantly higher than the normal group (16.0±2.3,13.2±4.3 respectively), the average grey scale values of ankle joints in CIA group 78±8 was significantly lower than the normal group 93±14 (P<0.05). Conclusion It has been found that MSCs can stay in the injured tissue and organs preferentially.

The homing of allogenic mesenchymal stem cells transplantation on rats with collagen induced arthritis

Objective To observe homing of mesenchymal stem cells (MSCs) in immune organs and inflammatory joints in collagen induced arthritis (CIA) rats. Methods Rats MSCs were isolated and expanded from bone marrow cells by density gradient centrifugation and adhering to the culture cell walls, and the phenotypes were assessed by flow cytometry. MSCs were labeled by PKH-26 and Brdu. Sixty-four rats were randomly divided into normal group and CIA group. Every 8 rats were sacrificed at 3, 11, 30, 42 days after transplantation of MSCs. At the end of the experiment, the specimens of thymus gland, spleen, ankle joints were exposed, fixed, decalcified, wrapped and cut into slices. Confocal laser scanning microscope and immunohistochemical method were used to observe migration and distribution of MSCs in different organs. Independent samples group t test with SPSS 12.0 software package was used for statistical analysis. Results It was found that allogenic MSCs could stay in spleen, thymus gland and joints of CIA rats for a relatively long period (42days). Forty-two days after transplantation of MSCs, the average grey scale values of spleen and thymus gland in CIA group (37.5±8.8, 29.9±5.9 respectively) were significantly higher than the normal group (16.0±2.3,13.2±4.3 respectively), the average grey scale values of ankle joints in CIA group 78±8 was significantly lower than the normal group 93±14 (P<0.05). Conclusion It has been found that MSCs can stay in the injured tissue and organs preferentially.