应用荧光原位杂交分析移植山羊体内的人源细胞  

Analysis of Human Cells in Transplanted Goats Using Fluorescence in Situ Hybridization

作　　者：肖艳萍[1] 陈美珏[1] 盛敏[1] 巩芷娟[1] 王舒[1] 黄淑帧[1] 

机构地区：[1]上海交通大学附属儿童医院,上海医学遗传研究所上海200040

出　　处：《中国医学科学院学报》2003年第2期129-133,共5页Acta Academiae Medicinae Sinicae

基　　金：国家"863"高技术项目(2002AA206211;2002AA216091)资助

摘　　要：目的 应用荧光原位杂交(fluorescence in situ hybridization,FISH)分析人造血干细胞(human hematopoietiestem cell,hHSC)经宫内移植至山羊体内后人源细胞的存在、比例及其动态变化。 方法 用人特异17号染色体卫星DNA探针对13头移植山羊进行间期FISH(interphase FISH,IFISH)分析,并用人男性特异Y染色体卫星DNA探针对其中2头移值人男性HSC的山羊进行IFISH分析。标本采用外周血细胞滴片、骨髓涂片和肝脏组织印片。结果 在13头移植山羊中,11头山羊体内有人源细胞存在,其中2头为人男性细胞,证实这些山羊为人/山羊异种移植嵌合体;人源细胞比例随山羊月龄递减,最长已存活21个月;所检测月龄段的移植山羊外周血、骨髓和肝脏组织中人源细胞比例小于1‰,但其中2头山羊肝脏组织中人源细胞集中一处,局部比例高达207.92‰和392.41‰。结论 FISH检测出人源细胞能在山羊体内存在并长期存活,表明山羊是经宫内移植hHSC的合适受体;人源细胞在外周血和骨髓中比例低,而在肝脏组织局部比例高,提示山羊肝脏组织局部内环境适于人源细胞存活、增殖和分化。Objective To analyze the existence and the dynamic cell frequencies of human cells in goats transplanted in utero with human hematopoietic stem cell (hHSC) by using fluorescence in situ hybridization (FISH) technique. Methods Interphase FISH (IFISH) with human-specific 17-chromosome satellite DNA and/or human-specific Y-chromosome satellite DNA as probes was performed to analyze the presence and proportions of human cells in 13 transplanted goats. Samples were peripheral blood cells, bone marrow smears and liver touch imprint preparations. Results Of the 13 transplanted goats, eleven were identified to present human cells. Among them, two goats transplanted with human male HSC were found to have human male cells. The results demonstrated that these transplanted goats were human/goal HSC xenogeneic chimeras. Human cell frequencies decreased with the goat age (months), but the longest survival reached 21 months. During the delected life periods of goats, human cell frequencies in peripheral blood, bone marrow and liver tissues were less than 1‰, but local human cell frequencies of 207.92‰ and 392.41‰ were detected in the liver tissues of 2 transplanted goats. Conclusions The existence and long-term survival of human cells in transplanted goats detected by FISH indicated that goats were appropriate recipients for hHSC in utero transplantation. The lower human cell frequencies in blood and bone marrow,and the higher local human cell frequencies in liver tissues suggested that the microenvi-ronment of goat liver tissues might favor the survival, proliferation and differentiation of human cells.

关 键 词：人造血于细胞 荧光原位杂交 移植 山羊 

分 类 号：Q26[生物学—细胞生物学] R329.24[医药卫生—人体解剖和组织胚胎学]