Yawata Y, Wada H, Kanzaki A, Yamada O
Department of Medicine, Kawasaki Medical School, Kurashiki, Japan.
Biomed Biochim Acta. 1990;49(2-3):S76-81.
A selective two phase liquid culture system for human erythroid progenitors was established, utilizing mononuclear cells obtained from human peripheral blood. The culture was performed in the presence of PHA-leukocyte conditioned medium for 7 days as the first phase, and nonphagocytic cells were recultured under hypoxic culture conditions containing iron-saturated transferrin and erythropoietin. During the second phase of the culture, differentiation to orthochromatic erythroblasts was observed on day 4, and to denucleated cells on day 8 with 16.8 +/- 0.7 micrograms/10(6) cells of hemoglobin level, respectively. Flow cytometric analyses revealed that, on day 3, cells became type M-positive, and the expression of ABH increased gradually corresponding to the extent of erythroid maturation. Lewis antigens and P antigens were expressed at day 5. The system is beneficial for studies on human membrane disorders, because the erythroid progenitors in human peripheral blood were utilized and the reasonable number of erythroid cells (0.5-1.5 x 10(7] was obtained with good erythroid maturation, such as denucleated erythrocytes.
利用从人外周血获得的单核细胞，建立了一种用于人类红系祖细胞的选择性两相液体培养系统。第一阶段，在PHA-白细胞条件培养基存在下进行培养7天，然后将非吞噬细胞在含有铁饱和转铁蛋白和促红细胞生成素的低氧培养条件下重新培养。在培养的第二阶段，第4天观察到向正染红细胞的分化，第8天观察到向无核细胞的分化，血红蛋白水平分别为16.8±0.7微克/10⁶个细胞。流式细胞术分析显示，第3天细胞变为M型阳性，ABH的表达随着红系成熟程度逐渐增加。第5天表达Lewis抗原和P抗原。该系统有利于人类膜疾病的研究，因为利用了人外周血中的红系祖细胞，并且获得了合理数量的红系细胞（0.5-1.5×10⁷），红系成熟良好，如无核红细胞。
