Xiao Yan-ping, Chen Mei-jue, Sheng Min, Gong Zhi-juan, Wang Shu, Huang Shu-zhen
Shanghai Institute of Medical Genetics, Children's Hospital, Shanghai Jiaolong University, Shanghai 200040, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2003 Apr;25(2):129-33.
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.
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.
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/goat HSC xenogeneic chimeras. Human cell frequencies decreased with the goat age (months), but the longest survival reached 21 months. During the detected life periods of goats, human cell frequencies in peripheral blood, bone marrow and liver tissues were less than 1@1000, but local human cell frequencies of 207.92@1000 and 392.41@1000 were detected in the liver tissues of 2 transplanted goats.
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 microenvironment of goat liver tissues might favor the survival, proliferation and differentiation of human cells.
采用荧光原位杂交(FISH)技术分析子宫内移植人造血干细胞(hHSC)的山羊体内人细胞的存在情况及动态细胞频率。
以人特异性17号染色体卫星DNA和/或人特异性Y染色体卫星DNA为探针进行间期FISH(IFISH),分析13只移植山羊体内人细胞的存在情况及比例。样本为外周血细胞、骨髓涂片和肝脏触片标本。
13只移植山羊中,11只被鉴定存在人细胞。其中,2只移植人男性HSC的山羊被发现含有人男性细胞。结果表明这些移植山羊是人/山羊HSC异种嵌合体。人细胞频率随山羊年龄(月)增加而降低,但最长存活时间达21个月。在检测的山羊存活期内,外周血、骨髓和肝脏组织中的人细胞频率低于1/1000,但在2只移植山羊的肝脏组织中检测到局部人细胞频率分别为207.92/1000和392.41/1000。
FISH检测显示移植山羊体内人细胞的存在及长期存活,表明山羊是子宫内hHSC移植的合适受体。血液和骨髓中人细胞频率较低,而肝脏组织中局部人细胞频率较高,提示山羊肝脏组织微环境可能有利于人细胞的存活、增殖和分化。