Drouet X, Douay L, Giarratana M C, Baillou C, Gorin N C, Salmon C, Najman A
Institut National de Transfusion Sanguine, Paris, France.
Br J Haematol. 1989 Oct;73(2):143-7. doi: 10.1111/j.1365-2141.1989.tb00243.x.
Prolonged in vitro maintenance of human bone marrow progenitor cells was achieved using a serum-free (SF) liquid culture system. Culture medium was based on Iscove's medium supplemented with bovine serum albumin, human transferrin, bovine insulin, soybean lecithin, cholesterol, hydrocortisone and alpha-thioglycerol. Under these standardized culture conditions, CFU-GM were maintained for up to 4 weeks, as is the case when using conventional serum-dependent medium. Erythropoiesis exhibited a slower decline than that found using serum containing medium. Development of normal haematopoiesis was effective in spite of poor stromal cell development--a confluent adherent layer as classically described in serum conditions was never achieved. Our newly defined system provides a reliable technique for studying human haematopoietic stem cell proliferation and differentiation in vitro; it allows for rational utilization of currently available purified recombinant growth factors. It may be a promising tool in the clinical use of cultured haematopoietic stem cells.
使用无血清(SF)液体培养系统实现了人骨髓祖细胞的长期体外维持。培养基基于Iscove培养基,并添加了牛血清白蛋白、人转铁蛋白、牛胰岛素、大豆卵磷脂、胆固醇、氢化可的松和α-硫甘油。在这些标准化培养条件下,CFU-GM最多可维持4周,这与使用传统血清依赖培养基的情况相同。红细胞生成的下降速度比使用含血清培养基时要慢。尽管基质细胞发育不良,但正常造血的发展仍然有效——从未达到血清条件下经典描述的汇合贴壁层。我们新定义的系统为体外研究人造血干细胞的增殖和分化提供了一种可靠的技术;它允许合理利用目前可用的纯化重组生长因子。它可能是培养造血干细胞临床应用中有前景的工具。