猕猴异体间充质干细胞输注安全性评价及异体存活分析

Safety evaluation of allogenic rhesus mesenchymal stem cells infusion and detection of chimerism

目的:探讨经静脉或骨髓内输注异体猕猴间充质干细胞(MSCs)的安全性以及异体MSCs是否能归巢骨髓并长期存活。方法:用密度梯度离心和贴壁筛选法分离、培养猕猴骨髓中的MSCs,观察其生物学特性。用流式细胞仪检测MSCs的细胞表型,采用不同培养条件诱导多向分化,用特异性染色鉴定;将雄性猕猴的MSCs经静脉输注或骨髓内输注给雌性猴,评价其不良反应和移植物抗宿主病(GVHD)表现,用PCR法检测受者骨髓中的Y特异性序列,评价MSCs在异体存活情况。结果:成功培养了猕猴的MSCs,多向分化实验证明其可分化为脂肪和成骨细胞;输注后未见急、慢性毒性反应和GVHD表现,部分雌性猕猴体内可检出Y特异性序列。结论:猕猴异体静脉或骨髓内输注MSCs未见急、慢性不良反应和GVHD,MSCs可在异体骨髓内存活。

Objective:To study the safety of intravenous (Ⅳ) and intra-bone marrow(IBM) infusion of allogenic rehsus mesenchymal stem cells (MSCs), and study the allogenic MSCs homing and living.Methods: MSCs were isolated and cultured; their biological characteristics were studied. The phenotypes of MSCs were detected by flow cytometry and the differentiated cells were identified with relevant specific staining. MSCs from male rhesus monkeys were infused into female rhesus monkeys via Ⅳ or IBM and the toxicity and graft-versus-host disease (GVHD) were evaluated. Y special sequence in recipients bone marrow was analyzed by polymerase chain reaction to evaluate the survival of MSCs.Results: Rhesus MSCs were successfully cultivated and they were capable of differentiating into lipocyte and osteoblast.No toxicity and GVHD were found by infusion of MSCs, and Y special sequence was detected in some recipient bone marrow.Conclusion: These data show that allogenic rhesus MSCs are not associated with significant toxicity and GVHD and are capable of establishing residence within the bone marrow following infusion via Ⅳ or IBM.