Zanjani E D, Pallavicini M G, Ascensao J L, Flake A W, Langlois R G, Reitsma M, MacKintosh F R, Stutes D, Harrison M R, Tavassoli M
Department of Veterans Affairs Medical Center, University of Nevada School of Medicine, Reno.
J Clin Invest. 1992 Apr;89(4):1178-88. doi: 10.1172/JCI115701.
Hemopoietic stem cells from human fetal liver were transplanted in utero into preimmune fetal sheep (48-54 days of gestation). The fate of donor cells was followed using karyotype analysis, by immunofluorescence labeling with anti-CD antibodies, and by fluorescent in situ hybridization using human-specific DNA probes. Engraftment occurred in 13 of 33 recipients. Of five live born sheep that exhibited chimerism, all expressed human cells in the marrow, whereas three expressed them in blood as well. Engraftment was multilineage (erythroid, myeloid, and lymphoid) and human hemopoietic progenitors (multipotent colony-forming units, colony-forming units-granulocyte, macrophage, and erythroid burst-forming units) capable of forming colonies in vitro were detected in all five lambs for greater than 2 yr. These progenitors responded to human-specific growth factors both in vitro and in vivo. Thus the administration of recombinant human IL-3 and granulocyte macrophage-colony-stimulating factor to chimeric sheep resulted in a 2.1-3.4-fold increase in the relative expression of donor (human) cells. These results demonstrate that the permissive environment of the preimmune fetal sheep provides suitable conditions for the engraftment and long-term multilineage expression of human hemopoietic stem cells in a large animal model. In this model, donor human cells appear to retain certain phenotypic and functional characteristics that can be used to manipulate the size of donor cell pool.
将来自人类胎儿肝脏的造血干细胞经子宫内移植到免疫前的胎羊（妊娠48 - 54天）体内。通过核型分析、使用抗CD抗体的免疫荧光标记以及使用人类特异性DNA探针的荧光原位杂交来追踪供体细胞的命运。33只受体中有13只发生了植入。在五只表现出嵌合现象的活产绵羊中，所有绵羊的骨髓中都表达人类细胞，而其中三只的血液中也表达人类细胞。植入是多谱系的（红细胞系、髓细胞系和淋巴细胞系），并且在所有五只羔羊中均检测到能够在体外形成集落的人类造血祖细胞（多能集落形成单位、粒细胞集落形成单位、巨噬细胞集落形成单位和红细胞爆式集落形成单位），持续时间超过2年。这些祖细胞在体外和体内均对人类特异性生长因子有反应。因此，向嵌合绵羊施用重组人白细胞介素-3和粒细胞巨噬细胞集落刺激因子导致供体（人类）细胞的相对表达增加了2.1 - 3.4倍。这些结果表明，免疫前胎羊的宽松环境为人类造血干细胞在大型动物模型中的植入和长期多谱系表达提供了适宜条件。在该模型中，供体人类细胞似乎保留了某些表型和功能特征，可用于操控供体细胞池的大小。
