人脐带间充质干细胞静脉输注小鼠的安全性

人脐带间充质干细胞静脉输注小鼠的安全性

何君1,2，李洋1,2，陈威1,2，郝好杰3，李名烁1,2，韩瑞红1,2，武鑫1,2，卢星辰1,2，金翠英1,2，郭珣1,2，易辉1,2，李晓岑1,2，赵颖1,2，武岩1,2，徐玉环1,2

1.中国医学科学院医学实验动物研究所，卫生部人类疾病比较医学重点实验室，北京，100021

2.中国医学科学院糖尿病研究中心，北京，100005

3.解放军总医院生命科学院基础医学所，北京，100853

【摘要】目的探讨C57/BL6J小鼠重复多次尾静脉输注人脐带间充质干细胞后的免疫反应和毒性。方法将SPF级别的32只C57/BL6J小鼠随机分为阴性对照组、细胞移植组，每组16 只，雌雄各半，细胞移植组小鼠尾静脉注射分离培养的第5代人脐带间充质干细胞，一次5×106/只，每周注射一次，连续注射4周；阴性对照组每次注射相同容积的PBS。注射后后观察小鼠的一般症状，末次注射后1周、4周进行血细胞计数、血生化、免疫反应指标、脏器质量测定和组织病理学检查。结果细胞移植组小鼠血细胞计数、血生化、脏器重量和脏器系数与对照组无显著性差异（P > 0.05），脏器组织病理学在光镜下检查结果与对照组无形态学差别，以及免疫结果测定（T细胞亚群CD3+、CD4+、CD8+及CD4+/CD8+）与对照组无显著性差异( P> 0.05)，较对照组有显著性差异（P 0.01）。结论人脐带间充质干细胞重复多次尾静脉输注C57/BL6J小鼠是安全可行的，对受者无明显免疫反应和毒副作用。【关键词】人脐带间充质干细胞；静脉输注；重复多次

【中图分类号】R332

A safety study of intravenous injection of human umbilical cord mesenchymal stem cells in mice

HE Jun1,2, LI Yang1,2, CHEN Wei1,2, HAO Hao-jie3, LI Ming-shuo1,2, HAN Rui-hong1,2, WU Xin1,2, LU Xing-chen1,2, JIN Cui-ying1,2, GUO Xun1,2, YI Hui1,2, LI Xiao-cen1,2, ZHAO Ying1,2, WU Yan1,2, XU Yu-huan1,2,

1.Institute of Laboratory Animal Science，Chinese Academy of Medical Science，Beijing 100021，China

2.Diabetes research center，Chinese Academy of Medical Science，Beijing 100005，China

3.Institute of Basic Medicine Science, College of Life Science, Chinese PLA General Hospital，Beijing 100853，China

[Abstract]Objective: To study the toxicity of intravenous injection of human umbilical cord mesenchymal stem cells in C57/BL6J mice with repeated administrations. Methods: 32 SPF C57/BL6J mice were randomly divided into negative control group and HUMSC transplantation group, of equal gender. HUMSCs were isolated and subsequently cultured through 5 passages in

vitro. 2×106 HUMSCs were injected each mice of transplantation group via tail vein once 1 week for 4 weeks, and the equal quantity PBS were injected to control group mice. Mice were euthanatized on the day of 1 week (8 mice per group) and 4 weeks(all survived mice) after the last treatment following the schedule. Toxicity was evaluated by clinical observations, pathology (blood cell counts, clinical biochemistry), immunologic consequences, visceral organs weight and anatomic pathology. Results:No significant difference in hematological and biochemical parameters between HUMSC transplantation group and control group (P> 0.05). There was also no significant change in organ weight or organ coefficient between the two groups(P > 0.05). There was no significant change in the subpopulation of T-cells (the percentage of CD3, CD4, CD8 T cells, and the ratio of CD4 and CD8 T cells). HUMSC transplantation group did not show any histopathological change in the viscera organs compared to control group. Conclusion:Results have suggested that intravenous injection of HUMSCs in C57/BL6J mice with repeated administrations is safe and feasible. No side effects and immune response were found in recipients.

[Key words]Human umbilical cord mesenchymal stem cell, Intravenous injection, Repeated administrations

【作者简介】何君（1969-），女，副研究员，博士，研究方向：药理学和毒理学

【基金项目】科技部863课题“干细胞治疗糖尿病的临床前方案优化及临床应用研究”

（2011AA020113和2013AA020105），十一五重大新药创制专项(2009ZX09303-008) 863 project of stem cells to treat diabetes clinical scheme optimization and clinical application research of the ministry of science and technology（2011AA020113 and 2013AA020105）, The 11th five-year fund of drug discovery projects(2009ZX09303-008)

脐带间充质干细胞（umbilical cord mesenchymal stem cells,UC-MSCs）具有多种作为种子细胞的优良特性，如在不同的诱导条件下可以向胰岛样细胞、肝样细胞、神经元样细胞、内皮细胞等进行分化[1-5]，连续传代培养和低温冻存后仍具有多向分化潜能，能造血支持和促进干细胞植入、免疫调控和自我复制，并且干细胞来源丰富、容易获得、增殖能力强,因此人脐带间充质干细胞被认为最具临床应用前途[6-11]。有文献报道将人脐带间充质干细胞移植大鼠和食蟹猴未见明显毒性反应，认为人脐带间充质干细胞异种移植是安全可行的[12-14]，但未见有移植到小鼠体内的安全性详细研究，因此本实验对C57/BL6J小鼠采用尾静脉多次重复输注人脐带间充质干细胞，通过检测输注后不同时间点小鼠的各项生化和免疫指标，探讨静脉输注脐带间充质干细胞的安全性，为开展人脐带间充质干细胞的临床应用提供参考，也为今后进行脐带间充质干细胞安全性评价和在体研究选择花费低、常规易得的实验动物提供初步的实验依据。