CYTOLOGICAL DEMONSTRATION OF CLONAL NATURE OF SPLEEN COLONIES DERIVED FROM TRANSPLANTED MOUSE MARROW CELLS
CYTOLOGICAL DEMONSTRATION OF CLONAL NATURE OF SPLEEN COLONIES DERIVED FROM TRANSPLANTED MOUSE MARROW CELLS
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DOI：
10.1038/197452a0
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发表时间：
1963-01-01
期刊：
影响因子：
64.8
通讯作者：
MCCULLOCH, EA
中科院分区：
文献类型：
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作者：
BECKER, AJ;TILL, JE;MCCULLOCH, EA
在正常的小鼠造血组织中，有一类细胞在移植到受到严重辐射的小鼠体内后，可以增殖并形成肉眼可见的集落。在脾脏中，以这种方式形成的集落是离散的，易于计数1，2。显微镜下，每个集落表现为一簇造血细胞，其中许多细胞正在分裂;通常，在给定的集落内，观察到的细胞表明分化沿沿着三条线发生，分别为红细胞、粒细胞和巨核细胞系列的细胞1.体外技术，其允许研究克隆群体，极大地推进了对细胞遗传和生理特性的认识。因此，需要证明是否每个脾集落都是由单个细胞发育而来的，因此是克隆;如果它们是，就有可能在体内研究造血细胞的克隆群体。支持集落是克隆这一观点的先前实验的证据如下：（1）移植的有核骨髓细胞数量与脾中发育的集落数量相关的曲线是线性的，并且没有显示初始阈值1，2;（2）可以形成集落的细胞的辐射存活曲线与细胞培养4或肿瘤移植5中单个细胞的存活曲线非常相似。这些观察结果只是间接地证明了这些菌落是克隆的。因此，试图获得菌落单细胞起源的直接证据似乎是可取的。乍一看，实现这一目标的最简单方法是利用具有已知染色体标记的骨髓细胞，如在T6/+小鼠中发现的骨髓细胞6，并将具有标记的骨髓细胞和不具有标记的骨髓细胞的混合物移植到受辐射的小鼠中。如果在脾脏中形成的集落总是由具有标记的细胞或不具有标记的细胞组成，那么当然，这表明单-
IN normal mouse hæmatopoietic tissue, there is a class of cells which, on being transplanted into heavily irradiated mice, can proliferate and form macroscopic colonies. In the spleen, the colonies formed in this manner are discrete and easy to count1, 2. Microscopically, each colony appears as a cluster of hæmatopoiotic cells, many of which are dividing; and often, within a given colony, the cells which are observed indicate that differentiation is occurring along three lines, into cells of the erythrocytic, granulocytic and megakaryocytic series, respectively1.In vitro techniques, which permit investigations of clonal populations, have greatly advanced the knowledge of the genetic and physiological properties of cells3. It was therefore desirable to prove whether or not spleen colonies each develop from single cells and hence are clones; for if they are, it would be possible to study clonal populations of hæmatopoietic cells in vivo. Evidence from previous experiments supporting the view that the colonies are clones is as follows:(1) the curve relating the number of nucleated marrow cells that are transplanted to the number of colonies that develop in the spleen is linear, and shows no initial threshold1, 2;(2) the radiation survival curve of cells that can form colonies closely resembles the survival curves obtained for single cells in cell culture4 or tumour transplants5. These observations provide only indirect evidence that the colonies are clones. It seemed desirable, therefore, to attempt to obtain direct evidence for the unicellular origin of the colonies. At first glance the easiest way to accomplish this aim would be to make use of marrow cells that have a known chromosome marker, such as that found in T6/+ mice6, and to transplant into irradiated mice a mixture of marrow cells with a marker and marrow cells with no marker. If the colonies that developed in the spleen were always composed of cells with markers or cells without markers it would, of course, suggest the single-