Miller J L, Njoroge J M, Gubin A N, Rodgers G P
Laboratory of Chemical Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Exp Hematol. 1999 Apr;27(4):624-9. doi: 10.1016/s0301-472x(98)00086-1.
We have developed a prospective approach to identify the generation of erythroid cells derived from cultured peripheral blood mononuclear cells (PBMC) by monitoring the expression of the cell surface protein CD48. Unpurified populations of PBMC obtained from the buffy coats of normal volunteers were grown in suspension culture in the absence or presence of erythropoietin. A profile of surface CD48 expression permitted a flow cytometric identification of erythropoietin responsive populations at various stages of their maturation. In the absence of erythropoietin (EPO) supplemented media, the CD48- cells represented <5% of the total population of PBMC remaining in culture. In cultures supplemented with 1 U/mL EPO, the mean percentage of CD48- cells increased to 34.7 + 14.9% (p < 0.01) after 14 days in culture. Coordinated CD34 and CD71 (transferrin receptor) expression, morphology, gamma-globin transcription, and colony formation in methylcellulose were observed during the 14-day culture period. Flow cytometric monitoring of bulk cultured PBMC provides a simple and reliable means for the prospective or real-time study of human erythropoiesis.
我们开发了一种前瞻性方法，通过监测细胞表面蛋白CD48的表达来鉴定源自培养的外周血单个核细胞（PBMC）的红系细胞生成。从正常志愿者的血沉棕黄层获得的未纯化PBMC群体在有无促红细胞生成素的情况下进行悬浮培养。表面CD48表达谱允许通过流式细胞术鉴定其成熟各个阶段对促红细胞生成素反应的群体。在没有补充促红细胞生成素（EPO）的培养基中，CD48-细胞占培养物中剩余PBMC总数的<5%。在补充有1 U/mL EPO的培养物中，培养14天后，CD48-细胞的平均百分比增加到34.7 + 14.9%（p < 0.01）。在14天的培养期内观察到CD34和CD71（转铁蛋白受体）表达、形态、γ-珠蛋白转录以及在甲基纤维素中的集落形成的协调情况。对大量培养的PBMC进行流式细胞术监测为人类红细胞生成的前瞻性或实时研究提供了一种简单可靠的方法。
