Kato K, Radbruch A
Institute for Genetics, University of Cologne, Germany.
Cytometry. 1993;14(4):384-92. doi: 10.1002/cyto.990140407.
CD34+ cells, present at a frequency of 0.18 +/- 0.052% among leukocytes from peripheral blood (PB), can be rapidly and efficiently (recovery of 39.0-74.0%) enriched to a frequency of 38.6-87.1% (54.4 +/- 12.3%) by high-gradient magnetic cell separation (MACS) for immunophenotyping, characterization in colony-forming cell (CFU) assays, and further purification to homogeneity (> 98%) by multiparameter fluorescence-activated cell sorting (FACS). Enriching PB-CD34+ cells for immunophenotyping allows the detection of small subpopulations, expressing the B-cell antigens CD10, CD19, and CD20, the T-cell antigens CD45RA and CD7, and a small subpopulation expressing high levels of CD34 (1.20 +/- 0.12%), which mostly coexpress CD19 (91.9 +/- 9.05%), CD20 (64.8 +/- 14.4%), and CD38 (84.5 +/- 10.3%). All PB-CD34+ cells express elevated levels of CD71 (transferrin receptor), with a subpopulation of high expressing cells, and CD38. Some cells express CD33. MACS-enriched PB-CD34+ cells show "normal" hematopoietic colony formation in vitro. The ease and efficiency of purification of large numbers of CD34+ cells from PB by MACS is not only relevant for the characterization of migrating stem cells but also opens new possibilities for stem cell transplantation and genetic manipulation of the hematopoietic system.
CD34+细胞在外周血（PB）白细胞中的出现频率为0.18±0.052%，通过高梯度磁性细胞分选（MACS）可快速有效地（回收率为39.0 - 74.0%）将其富集至频率为38.6 - 87.1%（54.4±12.3%），用于免疫表型分析、集落形成细胞（CFU）测定中的表征，以及通过多参数荧光激活细胞分选（FACS）进一步纯化至同质性（>98%）。富集PB - CD34+细胞用于免疫表型分析可检测到表达B细胞抗原CD10、CD19和CD20、T细胞抗原CD45RA和CD7的小亚群，以及表达高水平CD34（1.20±0.12%）的小亚群，其中大多数共表达CD19（91.9±9.05%）、CD20（64.8±14.4%）和CD38（84.5±10.3%）。所有PB - CD34+细胞均表达高水平的CD71（转铁蛋白受体），有一个高表达细胞亚群，以及CD38。一些细胞表达CD33。MACS富集的PB - CD34+细胞在体外显示出“正常”的造血集落形成。通过MACS从PB中纯化大量CD34+细胞的简便性和效率不仅与迁移干细胞的表征相关，也为造血系统的干细胞移植和基因操作开辟了新的可能性。
