DISPARATE DIFFERENTIATION IN MOUSE HEMATOPOIETIC COLONIES DERIVED FROM PAIRED PROGENITORS
DISPARATE DIFFERENTIATION IN MOUSE HEMATOPOIETIC COLONIES DERIVED FROM PAIRED PROGENITORS
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DOI：
10.1073/pnas.81.8.2520
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发表时间：
1984-01-01
期刊：
影响因子：
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通讯作者：
OGAWA, M
中科院分区：
文献类型：
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作者：
SUDA, T;SUDA, J;OGAWA, M
本文分析了配对祖细胞形成的小鼠造血集落在培养中的分化情况。采用显微操作技术从5-氟尿嘧啶处理小鼠脾脏培养的胚细胞菌落中分离单个祖细胞。18 ~ 24 h后，用微操作仪分离成对祖细胞，在含促红细胞生成素的甲基纤维素培养基和紫花草-有丝分裂原脾细胞条件培养基中培养。6 ~ 9 d后，分别摘取配对祖细胞的2个菌落，采用May-Grunwald-Giemsa染色进行差异计数。这里使用的缩写是n，中性粒细胞；米,巨噬细胞;e,嗜酸性粒细胞;桅杆，肥大细胞；米,巨核细胞;E,红细胞。在387对可评价的菌落中，有68对是由不同细胞系组合组成的菌落，如m- nmmmastem、m- nmmmastem、nm- nmmmastem、nmmastm - nmmastm、nmmast-nmmastM、nm-nmmastE、M-nmM、n-nmM、mM-nm、n-nmM、n-ne、m-mmast、m-mM、M-nm、E-nm、M-nm、M-m -m等。39对同源对显示相同的谱系组合，如nmmastEM、nmmastM、nmmast、mmastEM、nmEM、nme、nmM、mM和nm谱系。在其中一些对的成员中，单个细胞系的比例显着不同。其余的是一对单一血统的殖民地。从正常小鼠干细胞集落中获得的配对祖细胞也显示了同源和非同源细胞系的表达。将单个祖细胞衍生的群体表达量与单个祖细胞衍生的成对群体表达量的总和进行比较，表明其多样性不是由于微操作造成的损伤。实验数据支持干细胞分化的随机机制。
The differentiation of murine hemopoietic colonies derived from paired progenitors in culture was analyzed. Single progenitors were isolated by use of a micromanipulation technique from blast cell colonies cultured from the spleens of 5-fluorouracil-treated mice. Eighteen to 24 h later, the paired progenitors were separated with a micromanipulator and cultured in methylcellulose medium containing erythropoietin and pokeweed-mitogen spleen cell conditioned medium. Six to 9 days later, the 2 colonies derived from the paired progenitors were individually picked and differential counts were performed by using May-Grunwald-Giemsa stain. The abbreviations used here are n, neutrophil; m, macrophage; e, eosinophil; mast, mast cell; M, megakaryocyte; E, erythrocyte. Of a total of 387 pairs that could be evaluated, 68 were pairs of colonies consisting of dissimilar combinations of cell lineages such as m-nmmastEM, M-nmmastEM, nm-nmmastEM, nmmastM-nmmastEM, M-nmmastM, nmmast-nmmastM, nm-nmmastE, M-nmM, n-nmM, mM-nmM, m-nmmast, nm-nme, me-nm, mM-nm, n-ne, m-mmast, m-mM, M-nm, M-mM, E-nm, m-nm, M-m, etc. Thirty-nine were homologous pairs revealing identical lineage combinations such as nmmastEM, nmmastM, nmmast, mmastEM, nmEM, nme, nmM, mM and nm lineages. In members of some of these pairs, the proportions of the individual cell lineages were significantly different. The remainder were pairs of single lineage colonies. Paired progenitors obtained from the stem cell colonies of normal mice also revealed homologous and nonhomologous expression of the cell lineages. Comparison of lineage expression in colonies derived from single progenitors with the sum of lineages expressed in pairs of colonies derived from single progenitors indicated that the diversity was not due to injury inflicted by micromanipulation. Experimental data are provided in support of stochastic mechanisms of stem cell differentiation.