皮下移植羊膜间充质干细胞在正常小鼠体内的迁移及分化
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江苏省自然科学基金重点项目(BK2006729)


The migration and differentiation of β-gal transgenic mouse derived amniotic mesenchymal stem cells in recipient wild type mice after the transplantation subcutaneously
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    摘要:

    目的:探讨β-半乳糖苷酶(β-galactosidase,β-gal)转基因小鼠来源的羊膜间充质干细胞(amniotic mesenchymal stem cells,AMSCs)经皮下移植到野生型受体小鼠后向不同器官迁移分化情况-方法:从妊娠中晚期的β-gal转基因小鼠体内分离羊膜细胞,在含15% FBS的-琢-MEM完全培养基及成骨或成脂诱导培养基中培养-β-gal转基因小鼠来源的第2代AMSCs经皮下移植到新生C57BL/6野生型小鼠背部皮下,8周后,将受体小鼠各组织器官固定,Lac Z染色及石蜡包埋,并制成5 μm切片-切片经脱蜡,显微镜下观察拍照;或进行β-gal和胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)及β-gal和髓鞘碱性蛋白(myelin basic protein,MBP)免疫荧光双标染色-结果:β-gal转基因小鼠来源的第2代AMSCs呈现Lac Z染色阳性,经成骨或成脂培养基培养后可向成骨和脂肪细胞分化-β-gal转基因小鼠来源的第2代AMSCs植入野生型小鼠皮下,8周后,在野生型受体小鼠的甲状腺-肺-脾-脑内检测到Lac Z阳性的细胞-在受体小鼠脑中检测到β-gal和GFAP双阳性及β-gal和MBP双阳性的细胞-结论:皮下移植的AMSCs至少可以存活2个月,在正常小鼠体内经血液循环迁移到不同器官,并可分化成为不同器官的终末分化细胞,参与正常组织的自我更新,从而提示AMSCs可以作为干细胞移植的种子细胞,用于治疗不同疾病-

    Abstract:

    Objective:To assess the migration and differentiation capacity of β-galactosidase(β-gal) transgenic mouse derived amniotic mesenchymal stem cells (AMSCs) in recipient wild type mice after the transplantation subcutaneously. Methods:AMSCs were isolated from amnion membrane of the second-trimester β-gal transgenic mice and were cultured in either α-MEM containing 15% FBS, osteogenic- or adipogenic-induced medium. The second passage AMSCs derived from β-gal transgenic mice were transplanted subcutaneously into newborn C57BL/6 wild type mice. After 8 weeks,various organs from recipient mice were fixed and stained for Lac Z or embedded in paraffin. Then 5-μm sections were cut on a rotary microtome. The paraffin sections were dewax and observed under microscopy, and stained by immunofluorescence for β-gal,glial fibrillary acidic protein (GFAP) and myelin basic protein(MBP). Results:The second passage AMSCs derived from β-gal transgenic mice were positive for Lac Z staining and could differentiate into osteoblasts or adipocytes under cultures with the osteogenic- or adipogenic-induced medium. Followed subcutaneous injection of the second passage AMSCs derived from β-gal transgenic mice into wild type mice for 8 weeks,Lac Z positive cells were detected in thyroid,lung,spleen and brain of recipient wild type mice.The double positive cells for β-gal and GFAP, or β-gal and MBP, were detected in brain of recipient wild type mice. Conclusion:AMSCs transplanted through subcutaneously can survive at least 2 months,and migrate into multiorgans through circulation and differentiate into organ specific cells to participate in self-renewal of normal tissues. Our results indicate that AMSCs can be used as seed cells for stem cell therapy to treatment various diseases.

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谢春凤,苗登顺.皮下移植羊膜间充质干细胞在正常小鼠体内的迁移及分化[J].南京医科大学学报(自然科学版),2011,(5):601-605

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  • 收稿日期:2010-11-23
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