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[利用荧光原位杂交技术分析移植到山羊体内的人类细胞]

[Analysis of human cells in transplanted goats using fluorescence in situ hybridization].

作者信息

Xiao Yan-ping, Chen Mei-jue, Sheng Min, Gong Zhi-juan, Wang Shu, Huang Shu-zhen

机构信息

Shanghai Institute of Medical Genetics, Children's Hospital, Shanghai Jiaolong University, Shanghai 200040, China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2003 Apr;25(2):129-33.

PMID:12905704
Abstract

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/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.

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 microenvironment of goat liver tissues might favor the survival, proliferation and differentiation of human cells.

摘要

目的

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。

方法

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。

结果

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。

结论

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。

相似文献

1
[Analysis of human cells in transplanted goats using fluorescence in situ hybridization].[利用荧光原位杂交技术分析移植到山羊体内的人类细胞]
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2003 Apr;25(2):129-33.
2
[Detection of human/goat xenogeneic models by interphase fluorescence in situ hybridization].[通过间期荧光原位杂交检测人/山羊异种模型]
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2003 Apr;20(2):147-50.
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[Molecular identification of human/goat xenogeneic model].[人/山羊异种模型的分子鉴定]
Zhonghua Xue Ye Xue Za Zhi. 2002 Dec;23(12):634-7.
4
In utero transplantation of human hematopoetic stem cells into fetal goats under B-type ultrasonographic scan: an experimental model for the study of potential prenatal therapy.
Eur J Obstet Gynecol Reprod Biol. 2005 Feb 1;118(2):170-3. doi: 10.1016/j.ejogrb.2004.05.011.
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[The expression of human specific proteins in liver tissue of chimeric goats engrafted with human hematopoitic stem cells].[人造血干细胞移植嵌合体山羊肝脏组织中人类特异性蛋白质的表达]
Zhonghua Yi Xue Za Zhi. 2002 Jul;82(13):894-8.
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[A study of the engraftment, expansion and differentiation of human hematopoietic stem cells in goats].
Zhonghua Yi Xue Za Zhi. 2002 Jan 25;82(2):86-9.
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Hematopoietic stem cell transplantation in utero produces sheep-goat chimeras.子宫内造血干细胞移植可产生绵羊-山羊嵌合体。
Blood Cells Mol Dis. 2001 Jan-Feb;27(1):296-308. doi: 10.1006/bcmd.2000.0378.
8
Quantitative analysis of chimerism after allogeneic stem cell transplantation by real-time polymerase chain reaction with single nucleotide polymorphisms, standard tandem repeats, and Y-chromosome-specific sequences.通过使用单核苷酸多态性、标准串联重复序列和Y染色体特异性序列的实时聚合酶链反应对异基因干细胞移植后的嵌合现象进行定量分析。
Am J Hematol. 2006 Oct;81(10):735-46. doi: 10.1002/ajh.20693.
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Donor-derived type II pneumocytes are rare in the lungs of allogeneic hematopoietic cell transplant recipients.在异基因造血细胞移植受者的肺中,供体来源的II型肺泡上皮细胞很少见。
Ann Clin Lab Sci. 2006 Winter;36(1):47-52.
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In vivo formation of unstable heterokaryons after liver damage and hematopoietic stem cell/progenitor transplantation.肝损伤和造血干细胞/祖细胞移植后体内不稳定异核体的形成。
Stem Cells. 2006 Apr;24(4):1104-12. doi: 10.1634/stemcells.2005-0405. Epub 2005 Nov 10.

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Br J Haematol. 2014 Jul;166(2):268-78. doi: 10.1111/bjh.12870. Epub 2014 Mar 27.
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