CLONAL ORIGIN OF MURINE HEMATOPOIETIC COLONIES WITH APPARENT RESTRICTION TO GRANUCLOCYTE-MACROPHAGE-MEGAKARYOCYTE (GMM) DIFFERENTIATION
CLONAL ORIGIN OF MURINE HEMATOPOIETIC COLONIES WITH APPARENT RESTRICTION TO GRANUCLOCYTE-MACROPHAGE-MEGAKARYOCYTE (GMM) DIFFERENTIATION
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DOI：
10.1002/jcp.1041110304
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发表时间：
1982-01-01
影响因子：
5.6
通讯作者：
OGAWA, M
中科院分区：
文献类型：
--
作者：
NAKAHATA, T;OGAWA, M
表征了由粒细胞、巨噬细胞、巨核细胞和原始细胞组成且缺乏红系元素的小鼠造血集落。将小鼠骨髓或脾细胞在甲基纤维素培养基中在10%（体积/体积）商陆促分裂原刺激的脾细胞条件培养基（PWM-SCM）和2U/ml促红细胞生成素存在下培养8天。粒细胞-巨噬细胞-巨核细胞（GMM）集落与粒细胞-红细胞-巨噬细胞-巨核细胞（GEMM）集落不同，因为前者缺乏典型的红色爆发外观。在雄性和雌性骨髓细胞的混合实验中对Y染色体的分析证实了GMM集落的克隆性质。GMM集落的分类计数显示，在所有第8天和第12天的集落中以及在10个第14天的GMM集落中的7个中，原始细胞的数量不同但显著。胚细胞的百分比呈负相关的培养中的孵育时间的长度。再铺板实验证实，在所有50个第8天GMM集落中不存在晚期红系前体细胞，如CFU-E [红系集落形成单位]和正常成红细胞。然而，50个GMM集落中的6个含有能够红系表达的早期祖细胞，例如BFU-E [红细胞生成爆发形成细胞]、CFU-GEM [红系巨噬细胞集落形成单位]、CFU-GEM [粒细胞-红细胞-巨噬细胞集落形成单位]和CFU-GEMM [粒细胞-红细胞-巨噬细胞-巨核细胞集落形成单位]。相比之下，未显示原始细胞的3天-14 GMM集落不能产生次级集落。虽然后者集落的祖细胞仅限于粒细胞-巨噬细胞-巨核细胞分化，但一些含有母细胞的明显GMM集落可能起源于接近多能干细胞的早期祖细胞。详细的细胞学分析和重新铺板实验是必要的混合集落在文化的真正分化潜力的表征。
Murine hemopoietic colonies consisting of granulocytes, macrophages, megakaryocytes and blast cells and lacking erythroid elements were characterized. Mouse marrow or spleen cells were cultured in methylcellulose media in the presence of 10% (vol/vol) pokeweek mitogen-stimulated spleen cell-conditioned medium (PWM-SCM) and 2 U/ml erythropoietin for 8 days. Granulocyte-macrophage-megakaryocyte (GMM) colonies were distinguished from granulocyte-erythrocyte-macrophage-megakaryocyte (GEMM) colonies because the former lacked the typical appearance of bursts with red color. Analysis of Y-chromosomes in mixing experiments with male and female marrow cells confirmed the clonal nature of the GMM colonies. Differential counts of GMM colonies revealed varying, but significant, numbers of blast cells in all of the day-8 and day-12 colonies and in 7 of 10 day-14 GMM colonies. The percentages of blast cells were inversely related to the length of incubation in culture. Replating experiments confirmed the absence of late erythroid precursors such as CFU-E [erythroid colony-forming units] and normoblasts in all of the 50 day-8 GMM colonies. However, 6 of the 50 GMM colonies contained early progenitors capable of erythroid expression, such as BFU-E [erythropoietic burst-forming cell], CFU-GEM [erythroid macrophage colony-forming unit], CFU-GEM [granulocyte-erythrocyte-macrophage colony-forming unit] and CFU-GEMM [granulocyte erythrocyte-macrophage-megakaryocyte colony-forming unit]. In contrast, the 3 day-14 GMM colonies which did not reveal blast cells failed to produce secondary colonies. While the progenitors for the latter colonies are restricted to only granulocyte-macrophage-megakaryocyte differentiation, some of the apparent GMM colonies containing blast cells may have originated in early progenitors close to pluripotent stem cells. Detailed cytological analyses and replating experiments are necessary for characterization of true differentiation potentials of mixed colonies in culture.