异基因造血干细胞移植后受者骨髓的间充质干细胞来源研究_医学论文.docVIP

异基因造血干细胞移植后受者骨髓的间充质干细胞来源研究_医学论文 异基因造血干细胞移植后受者骨髓的间充质干细胞来源研究_医学论文 【摘要】 为了研究造血干细胞移植植活患者骨髓源间充质干细胞（MSC）的来源，选择临床上供受者性别不同的造血干细胞移植后临床植活的34例患者作为研究对象，抽取骨髓，进行间充质干细胞培养传代，利用流式细胞术 (FCM) 检测患者MSC的表面抗原表达特征，利用聚合酶链反应 (PCR) 分析患者MSC的釉基质基因表达情况以确定其性别来源，利用荧光原位杂交 (FISH) 技术进一步证实患者MSC的性别来源。结果表明：34例患者中有31例患者的MSC原代培养能达到融合，其中24例患者成功传代5代以上，且可进行PCR及FISH检测；FCM结果显示，第5代MSC的CD14+CD45+细胞表达低于0.04%；PCR结果显示，移植后患者的MSC源于患者本人；FISH结果显示，移植后患者的MSC 100%源于患者本人。结论：造血干细胞移植后患者骨髓中的MSC仍源于患者本人。 【关键词】 造血干细胞移植 　　Origin of Mesenchymal Stem Cells in Bone Marrow of Patients after Allogeneic Stem Cell Transplantation 　　Abstract The aim of this study was to investigate the origin of bone marrow-derived mesenchymal cells in 34 patients who had received a sex-mismatched hematopoietic stem cells transplant (HSCT).The mesenchymal stem cells (MSC) from 34 patients were collected for test.The different passage MSC of patients were analyzed by flow cytometry (FCM) to reveal the cell surface antigen expression.The polymerase chain reaction (PCR) analysis of the amelogenin (AMEL) genes was used to detect donor cells from host cells.The cultured MSC were stained by fluorescence in situ hybridization (FISH) probes for chromosomes X (Xp11.1-Xq11.1) and Y (Yq12) to distinguish donor cells from host cells.The results indicated that 31out of 34 cases showed the confluent stroma and 24 out of these 31 cases were successfully passaged for more than 5 generations which could be used for PCR and FISH analysis.According to FCM results the number of CD14+CD45+ cells，which was regarded as monocyte/macrophage from the cultured MSC (passage 5) was less than 0.04%.In PCR assay，the marrow-derived MSC from the passage 5 were found to be of host origin. FISH assay demonstrated that marrow-derived MSC from the passage 5 were found to be of host origin up to 100%.It is concluded that the origin of MSC in bone marrow of patients after allogeneic stem cell transplantation was confirmed to be derived still from the recipients their own. 　　Key words hemotopo