Huang S, Terstappen L W
Becton Dickinson Immunocytometry Systems, San Jose, CA 95131.
Blood. 1994 Mar 15;83(6):1515-26.
Multilineage differentiation of human fetal bone marrow CD34+ cell subsets was examined using a single-cell liquid culture assay. Four CD34+ cell populations, ie, (1) CD38-, HLA-DR+, (2) CD38-, HLA-DR-, (3) CD38+, HLA-DR-, and (4) CD38+, HLA-DR+ cells, were sorted as single cells into 96-well flat-bottom culture plates containing long-term culture medium supplemented with interleukin-3, interleukin-6, stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor, erythropoietin, basic fibroblast growth factor (bFGF), and insulin-like growth factor-1 (IGF-1). Single CD34+, CD38-, HLA-DR+ cells had the highest replating efficiency as well as the highest replating efficiency. The cellular composition of the single-cell progeny was studied by morphologic and/or flow cytometric examination. Only the progeny of single CD34+ cells that lacked CD38 could give rise to each of the hematopoietic cell lineages. The expansion of the progeny of single CD34+, CD38-, HLA-DR+ cells was examined in more detail and showed three clearly distinguishable growth patterns: 28% (SD, +/- 10%; n = 14) of the single cells formed cell clusters/colonies; 9% (SD, +/- 4%; n = 14) formed dispersed cells; and 11% (SD, +/- 6%; n = 14) gave rise to a mixture of cell clusters and dispersed cells. The dispersed cell growth pattern was reduced when SCF or bFGF and IGF-1 was absent in the growth factor cocktail. The replating ability of the dispersed cells was considerably larger than that of cells with other growth patterns, in that 76% of the cells that gave rise to dispersed cells and 54% of the cells that gave rise to dispersed cells as well as cell clusters gave rise to a second generation, but only 7% of the cells that gave rise to cell clusters gave rise to a second generation. The second generation of cells continued to produce third and fourth generations after repetitive replating, except for the replated cells from cell clusters. In contrast with the first-generation progeny, SCF did not have an influence on the replating ability of the cells. Only in the progeny of single CD34+, CD38-, HLA-DR+ cells that gave rise to dispersed cells was each of the hematopoietic cell lineages found, ie, B lymphocytes, neutrophils, monocytes, macrophages, osteoclasts, basophils/mast cells, eosinophils, erythrocytes, megakaryocytes, and platelets.(ABSTRACT TRUNCATED AT 400 WORDS)
采用单细胞液体培养试验检测人胎儿骨髓CD34+细胞亚群的多谱系分化。将四个CD34+细胞群体，即(1) CD38-、HLA-DR+，(2) CD38-、HLA-DR-，(3) CD38+、HLA-DR-，以及(4) CD38+、HLA-DR+细胞，作为单个细胞分选到含有补充了白细胞介素-3、白细胞介素-6、干细胞因子(SCF)、粒细胞-巨噬细胞集落刺激因子、促红细胞生成素、碱性成纤维细胞生长因子(bFGF)和胰岛素样生长因子-1(IGF-1)的长期培养基的96孔平底培养板中。单个CD34+、CD38-、HLA-DR+细胞具有最高的再接种效率。通过形态学和/或流式细胞术检查研究单细胞后代的细胞组成。只有缺乏CD38的单个CD34+细胞的后代能够产生各造血细胞谱系。对单个CD34+、CD38-、HLA-DR+细胞后代的扩增进行了更详细的检查，显示出三种明显可区分的生长模式：28%(标准差，±10%；n = 14)的单个细胞形成细胞簇/集落；9%(标准差，±4%；n = 14)形成分散细胞；11%(标准差，±6%；n = 14)产生细胞簇和分散细胞的混合物。当生长因子混合物中不存在SCF或bFGF和IGF-1时，分散细胞生长模式减少。分散细胞的再接种能力明显大于具有其他生长模式的细胞，即产生分散细胞的细胞中有76%以及产生分散细胞和细胞簇的细胞中有54%产生第二代，但产生细胞簇的细胞中只有7%产生第二代。除了来自细胞簇的再接种细胞外，第二代细胞在重复再接种后继续产生第三代和第四代。与第一代后代不同，SCF对细胞的再接种能力没有影响。只有在产生分散细胞的单个CD34+、CD38-、HLA-DR+细胞的后代中才发现各造血细胞谱系，即B淋巴细胞、中性粒细胞、单核细胞、巨噬细胞、破骨细胞、嗜碱性粒细胞/肥大细胞、嗜酸性粒细胞、红细胞、巨核细胞和血小板。(摘要截断于400字)
