Hematopoietic stem cell engraftment by early-stage in utero transplantation in a mouse model

被引:12
|
作者
Chen, Xin [1 ,3 ]
Gong, Xiu-li [1 ,3 ]
Katsumata, Makoto [1 ,4 ]
Zeng, Yi-tao [1 ,3 ]
Huang, Shu-zhen [1 ,3 ]
Zeng, Fanyi [1 ,2 ,3 ]
机构
[1] Shanghai Jiao Tong Univ, Shanghai Childrens Hosp, Shanghai Inst Med Genet, Sch Med, Shanghai 200040, Peoples R China
[2] Shanghai Jiao Tong Univ, Sch Med, Inst Med Sci, Shanghai 200040, Peoples R China
[3] Minist Hlth China, Key Lab Embryo Mol Biol, Shanghai, Peoples R China
[4] Univ Penn, Sch Med, Dept Pathol & Lab Med, Philadelphia, PA 19104 USA
关键词
In utero transplantation; Hematopoietic stem cell; Chimera establishment; Allogeneic transplantation; Fluorescence in situ hybridization; GENE-EXPRESSION; CHIMERISM; MICE;
D O I
10.1016/j.yexmp.2009.07.009
中图分类号
R36 [病理学];
学科分类号
100104 ;
摘要
A novel intrauterine transplantation (IUT) approach was developed to improve the efficiency of engraftment of hematopoietic stem cells (HSCs). HSCs with a green fluorescent protein (GFP) reporter gene were transplanted in utero on days 12.5,13.5 and 14.5 post coitum (p.c.). The degree of chimerism of donor cells in recipient newborn mice was examined using fluorescent microscopy, polymerase chain reaction (PCR), fluorescence-activated cell sorting (FACS), and fluorescence in situ hybridization (FISH) analyses. Microscopic examination revealed the presence of green fluorescent signal in the peripheral blood of the chimeric mice. The highest survival rate (47%) as well as the highest chimerism rate (73%) were achieved by our new approach in the newborn mice that were subjected to in utero transplantation (IUT) on day 12.5 p.c. (E12.5) compared to the conventional IUT method. FACS analysis indicated that 1.55 +/- 1.10% of peripheral blood cells from the newborn mice were GFP-positive donor cells. FISH showed that cells containing the donor-specific GFP sequence were present in the bone marrow (BM) of the chimeric mice. Thus, the efficiency of chimera production with this new method of IUT was significantly improved over the existing IUT techniques and instruments. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码:173 / 177
页数:5
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