Mouchiroud G, Berthier R, Newton I A, Chapel A, Hollard D
Exp Hematol. 1985 Jul;13(6):566-73.
Monoclonal antibodies against myeloid cell surface antigens were selected according to their reactivity against normal human bone marrow cells. Each of the antibodies detected a different population of maturing bone marrow cells, but almost none of the progenitor cells assayed (seven-day GM-CFC, 14-day GM-CFC, CFU-E, BFU-E). Myeloid cells, erythroid cells, and the majority of lymphocytes and monocytes were simultaneously depleted with the mixture of these three antibodies using fluorescence-activated cell sorting (FACS) or panning for cell separation; 14-day GM-CFC and BFU-E were enriched six- to 16-fold when negative cells were sorted. After a negative selection step using panning, BFU-E and 14-day GM-CFC were enriched four- to seven-fold and five- to 13-fold, respectively. Negative cells obtained after panning or cell sorting were also enriched in blast cells (24.4% and 27.2%, respectively) and depleted in maturing bone marrow cells, with the exception of variable numbers of lymphocytes, monocytes, and plasma cells. FACS of negative cells using both forward light-scatter and perpendicular light-scatter parameters resulted in a cell population that contained a majority of undifferentiated blasts and 5.5%-9.6% BFU-E and 14-day GM-CFC.
根据抗正常人骨髓细胞的反应性，筛选出针对髓样细胞表面抗原的单克隆抗体。每种抗体都能检测到不同群体的成熟骨髓细胞，但几乎检测不到所检测的祖细胞（7天粒-巨噬细胞集落形成细胞、14天粒-巨噬细胞集落形成细胞、红细胞集落形成单位、爆式红细胞集落形成单位）。使用荧光激活细胞分选（FACS）或淘选法进行细胞分离时，这三种抗体的混合物可同时清除髓样细胞、红系细胞以及大多数淋巴细胞和单核细胞；当分选阴性细胞时，14天粒-巨噬细胞集落形成细胞和爆式红细胞集落形成单位可富集6至16倍。经过淘选的阴性选择步骤后，爆式红细胞集落形成单位和14天粒-巨噬细胞集落形成细胞分别富集4至7倍和5至13倍。淘选或细胞分选后获得的阴性细胞中也富含原始细胞（分别为24.4%和27.2%），成熟骨髓细胞减少，但有数量不等的淋巴细胞、单核细胞和浆细胞除外。使用前向光散射和垂直光散射参数对阴性细胞进行FACS分析，得到的细胞群体中大多数是未分化的原始细胞，还有5.5%-9.6%的爆式红细胞集落形成单位和14天粒-巨噬细胞集落形成单位。
