Preisler H D, Epstein J
J Lab Clin Med. 1979 Sep;94(3):414-20.
A simple method has been developed for obtaining specimens of human marrow which are enriched for myeloblasts and promyelocytes. The erythrocytes are lysed, the marrow is incubated with iron particles, and the cells that phagocytize the iron are removed with a powerful magnet. The marrow is then subjected to a density-cut centrifugation using Ficol-Hypaque with a density of 1.084 gm/mm3. The cells that do not enter the Ficol-Hypaque are removed from the surface and studied. The proportion of myeloblasts and promyelocytes in this subpopulation of cells exceeds 50%. Total recovery of these immature myeloid progenitor cells is 50% of that in the original marrow specimen. This method has been used for cell suspensions containing as many as 10(9) cells. Cells prepared using this method incorporate 3H-TdR, 3H-UR, and 3H-Leu at a higher rate than the unseparated specimens and have a cloning efficiency of 2.0% to 19.4% compared with 0.1% to 2.17% for the unseparated marrows.
已开发出一种简单方法来获取富含成髓细胞和早幼粒细胞的人类骨髓标本。红细胞被裂解,骨髓与铁颗粒一起孵育,然后用强力磁铁去除吞噬铁的细胞。接着使用密度为1.084克/立方毫米的Ficoll-泛影葡胺对骨髓进行密度梯度离心。未进入Ficoll-泛影葡胺的细胞从表面去除并进行研究。该细胞亚群中,成髓细胞和早幼粒细胞的比例超过50%。这些未成熟髓系祖细胞的总回收率为原始骨髓标本的50%。此方法已用于多达10⁹个细胞的细胞悬液。用此方法制备的细胞比未分离的标本更高效地掺入³H-TdR、³H-UR和³H-Leu,克隆效率为2.0%至19.4%,而未分离的骨髓克隆效率为0.1%至2.17%。