Kirshner J J, Goldberg J
Exp Hematol. 1980 Nov;8(10):1202-7.
Four types of colonies can be formed by progenitor cells (CFUc) in double layer agar cultures: eosinophil, granulocyte, macrophage, and mixed granulocyte-macrophage. We have developed a method for in situ staining of intact agar plates with luxol fast blue and acetoorcein employed to differentiate the type of colonies. Cultures of 10(5) normal human bone marrow cells yielded 89.1 colonies, of which 72% were eosinophil. Cultures of 5 X 10(5) peripheral blood cells, enriched by isopycnic sedimentation, yielded 39.0 colonies, of which 48% were eosinophil. T-cell depleted peripheral blood mononuclear cells yielded 116.9 colonies/5 X 10(5) cells, of which 74% were eosinophil. Umbilical cord blood cells yielded 65.1 colonies/5 X 10(5) cells, of which 23% were eosinophil. Comparing similar numbers of total colonies, there appears to be fewer relative and absolute numbers of eosinophil colonies formed by CFUc recovered from cord blood as opposed to adult peripheral blood and bone marrow. Enumeration of colony numbers without regard to types of colonies is imprecise. The method employed in the present study is easy to perform, reproducible and provides permanent slides.
祖细胞(CFUc)在双层琼脂培养中可形成四种类型的集落:嗜酸性粒细胞集落、粒细胞集落、巨噬细胞集落以及粒细胞 - 巨噬细胞混合集落。我们已开发出一种方法,用卢戈氏坚牢蓝和醋地衣红对完整的琼脂平板进行原位染色,以区分集落类型。10⁵个正常人骨髓细胞培养物产生了89.1个集落,其中72%为嗜酸性粒细胞集落。通过等密度沉降富集的5×10⁵个外周血细胞培养物产生了39.0个集落,其中48%为嗜酸性粒细胞集落。去除T细胞的外周血单个核细胞产生116.9个集落/5×10⁵个细胞,其中74%为嗜酸性粒细胞集落。脐带血细胞产生65.1个集落/5×10⁵个细胞,其中23%为嗜酸性粒细胞集落。比较相似数量的总集落数,与成人外周血和骨髓相比,从脐带血中回收的CFUc形成的嗜酸性粒细胞集落的相对数量和绝对数量似乎较少。不考虑集落类型而对集落数量进行计数是不精确的。本研究中采用的方法易于操作、可重复且能提供永久切片。