Establishment of Human/Mouse Chimerism by in Utero Transplantation of Human Umbilical Cord Blood CD34(superscript +)Lin(superscript -) Cells into Fetal Mice

Yu-Dan Fang ； Zhi-Juan Gong ； Mei-Jue Chen ； Min Cheng ； Jing Fu ； Shu-Zhen Huang

《Journal of Genetics and Molecular Biology》 17卷2期 (2006/06) Pp. 94-103

https://doi.org/10.30047/JGMB.200606.0003

並列摘要

To investigate the biological behavior of the human hematopoetic stein cells (HSC) in vivo, we have established a human/mouse xenogeneic transplant chemiric animal model by in utero transplantation of human HSC/progenitor cells into fetal mice. Moreover, the long-term survival of the human cells in the transplant mice was studied. The human CD34(superscript +)lin(superscript -) cells derived from umbilical cord blood were injected into the peritoneal cavity of the fetal mice at 12.5-13 days of gestation, and a variety of methods, including fluorescent activated cell sorting (FAGS) analysis, nested-PCR, real-time PCR, reverse transcription (RT)-PCR as well as immunohistochemical assas were used to test the features for the recipient mice after birth at intervals. Of 40 live-born recipients, 12 had the presence of human CD34(superscript +) and GPA+ cells in peripheral blood by FAGS assessment. Real-time quantitative PCR detection showed that the chimeric dose of donor cells in peripheral blood and livers of the grafted recipients ranged from 1/2000 to 1/30. RT-PCR analysis showed that human CD34(superscript +) and GPA(superscript +) mRNAs were identified in peripheral blood and livers in three tested recipients. Significantly, human serum albumin (hALB) mRNA also expressed in the livers of these mice. Simultaneously, immunohistochemical analysis was in accordance with the above results. All these results indicated that the human cells engrafted in the mice can be survived and hold biological activity. This human/mouse chimerism model, therefore, can provide an effective tool for the studies of the biological behavior of the human HSC in vivo and the potential implication of prenatal treatment.

並列關鍵字

human hematopoietic stem cell hHSC ； xenogeneic transplantation ； in utero ； mouse ； chimerism