Spence S E, Keller J R, Ruscetti F W, Czarra K T, Gooya J M, Funakoshi S, Longo D L, Murphy W J
Biological Carcinogenesis and Development Program, Program Resources, Inc/DynCorp, National Cancer Institute, Frederick Cancer Research and Development Center, MD, USA.
Leukemia. 1995 Oct;9 Suppl 1:S43-7.
Demonstration of the ability of fresh human hematopoietic cells to engraft severe combined immuno-deficient (scid) mice has provided an in vivo assay for expansion and maturation of early human progenitor cells. However, engraftment of cultured hematopoietic cells has been difficult to achieve. We wished to further develop this model as an in vivo assay for efficiency of retroviral gene transfer and expression in the differentiated progeny of adult human bone marrow progenitor cells. Human bone marrow cells were cultured in vitro for six days under conditions suitable for infection by retroviral vectors prior to transfer to irradiated scid mice. Cultured human bone marrow cells introduced by both intravenous (i.v.) and intraperitoneal (i.p.) injection persisted in the bone marrow, spleen and peritoneum of recipient animals up to four weeks after transfer. Following irradiation scid mice receiving cultured human bone marrow cells by either i.v. or i.p. routes demonstrated engraftment of the bone marrow and spleen as determined by the growth of human hematopoietic progenitors in soft agar. By flow cytometric analysis human cells were also detected in the peritoneum of mice receiving cultured human bone marrow cells i.p. These results suggest that the transfer of cultured human bone marrow cells to scid mice with the subsequent engraftment of these cells in the bone marrow, spleen and peritoneum of recipients can routinely occur. This provides an in vivo model for retroviral gene transfer to human cells.
新鲜人类造血细胞植入严重联合免疫缺陷（scid）小鼠的能力证明，为早期人类祖细胞的扩增和成熟提供了一种体内检测方法。然而，培养的造血细胞的植入一直难以实现。我们希望进一步开发这个模型，作为一种体内检测方法，用于检测逆转录病毒基因在成人骨髓祖细胞分化后代中的转移和表达效率。在转移到经辐照的scid小鼠之前，将人类骨髓细胞在适合逆转录病毒载体感染的条件下体外培养6天。通过静脉内（i.v.）和腹腔内（i.p.）注射引入的培养人类骨髓细胞在转移后长达四周的时间里持续存在于受体动物的骨髓、脾脏和腹膜中。在接受经静脉或腹腔途径注入培养人类骨髓细胞的辐照scid小鼠中，通过软琼脂中人类造血祖细胞的生长确定骨髓和脾脏出现了植入。通过流式细胞术分析，在接受腹腔内注入培养人类骨髓细胞的小鼠腹膜中也检测到了人类细胞。这些结果表明，将培养的人类骨髓细胞转移到scid小鼠中，随后这些细胞在受体的骨髓、脾脏和腹膜中植入可以常规发生。这为逆转录病毒基因转移到人类细胞提供了一种体内模型。
