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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 stemcells,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α-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5μm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 basicprotein,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。
AIM: To investigate the transdifferentiation of amniotic mesenchymal stem cells (AMSCs) derived from β-galactosidase (β-gal) transgenic mice into different organs after subcutaneous transplantation into wild-type mice Differentiation. Methods: Amniotic cells were isolated from β-gal transgenic mice in the third trimester of pregnancy and cultured in α-MEM complete medium containing 15% FBS and osteogenic or adipogenic medium. Generation 2 AMSCs from β-gal transgenic mice were subcutaneously transplanted subcutaneously into the dorsal skin of newborn C57BL / 6 wild-type mice. After 8 weeks, the tissues and organs of the recipient mice were fixed, Lac Z staining and paraffin-embedded 5 μm sections were made. Sections were deparaffinized and photographed under a microscope; or double-stained with β-gal and glial fibrillary acidic protein (GFAP) and β-gal and myelin basic protein (MBP) . Results: The second generation of AMSCs derived from β-gal transgenic mice showed positive Lac Z staining and could differentiate into osteoblasts and adipocytes after osteogenic or adipogenic culture. Generation 2 AMSCs from β-gal transgenic mice were subcutaneously implanted into wild-type mice. After 8 weeks, Lac Z-positive cells were detected in the thyroid, lung, spleen and brain of wild-type recipient mice. Both β-gal and GFAP double positive and β-gal and MBP double positive cells were detected in the recipient mouse brain. CONCLUSIONS: AMSCs transplanted subcutaneously can survive for at least 2 months and migrate to different organs through blood circulation in normal mice and can differentiate into terminal differentiated cells of different organs and participate in the self-renewal of normal tissues, thus suggesting that AMSCs can act as Stem cell transplanted seed cells for the treatment of different diseases.