Zanjani E D, Flake A W, Rice H, Hedrick M, Tavassoli M
Department of Medicine, Department of Veterans Affairs Medical Center, University of Nevada School of Medicine, Reno 89520.
J Clin Invest. 1994 Mar;93(3):1051-5. doi: 10.1172/JCI117054.
We previously reported on the successful engraftment and long-term multilineage expression (erythroid, myeloid, lymphoid) of human fetal liver hematopoietic stem cells in sheep after transplantation in utero. That the engraftment of long-term repopulating pluripotent stem cells occurred in these animals was shown here by the fact that transplantation of human CD45+ cells isolated from bone marrow of these chimeric animals into preimmune fetal sheep resulted in engraftment and expression of human cells. Marrow cells were obtained from three chimeric sheep at 3.2-3.6 yr after transplant. The relative percentage of human CD45+ cells present in these marrows was 3.3 +/- 0.32%. A total of 29 x 10(6) CD45+ cells were isolated by panning, pooled, and transplanted into six preimmune sheep fetuses (4.8 x 10(6) cells/fetus). All six recipients were born alive. Hematopoietic progenitors exhibiting human karyotype were detected in marrows of two lambs soon after birth. Cells expressing human CD45 antigen were also detected in blood and marrow of both lambs. Human cell expression has been multilineage and has persisted for > 1 yr. These results demonstrate that the expression of human cells in this large animal model resulted from engraftment of long-term repopulating pluripotent human stem cells.
我们之前报道了人胎肝造血干细胞在子宫内移植到绵羊体内后成功植入并长期进行多系表达(红细胞系、髓细胞系、淋巴细胞系)的情况。这些动物体内发生了长期重建造血的多能干细胞植入,这一点通过以下事实得以证明:将从这些嵌合动物骨髓中分离出的人CD45+细胞移植到未免疫的胎羊体内,可导致人细胞的植入和表达。在移植后3.2 - 3.6年从三只嵌合绵羊获取骨髓细胞。这些骨髓中人CD45+细胞的相对百分比为3.3±0.32%。通过淘选共分离出29×10⁶个CD45+细胞,汇集后移植到六只未免疫的胎羊(4.8×10⁶个细胞/胎羊)体内。所有六只受体均存活出生。出生后不久在两只羔羊的骨髓中检测到呈现人核型的造血祖细胞。在两只羔羊的血液和骨髓中也检测到表达人CD45抗原的细胞。人细胞表达具有多系性且持续超过1年。这些结果表明,在这个大型动物模型中人细胞的表达是由长期重建造血的多能人类干细胞植入所致。