CLONAL APPROACH TO PROBLEM OF NEURAL CREST DETERMINATION
CLONAL APPROACH TO PROBLEM OF NEURAL CREST DETERMINATION
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DOI：
10.1016/0012-1606(75)90104-9
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发表时间：
1975-01-01
影响因子：
2.7
通讯作者：
KONIGSBERG, IR
中科院分区：
文献类型：
--
作者：
COHEN, AM;KONIGSBERG, IR
关于神经嵴发育的一个基本问题是这种胚胎组织可能的多能性。作为研究这个问题的第一步，克隆技术用于产生嵴细胞的同质群体。这些细胞的原代培养是通过体外分离日本鹌鹑的神经管并使嵴细胞迁移而获得的。取出外植体，分离生长物，用胰蛋白酶解离，并将细胞以克隆密度铺板。以这种方式衍生的集落分为以下几类：殖民地的所有细胞都着色;没有细胞着色;以及一些细胞着色，其余的未着色。着色集落通常来自小的圆形细胞，而非着色集落通常来自大的扁平多形细胞。黑素细胞的分化并不妨碍其继续增殖。此外，色素表型在至少25代中是稳定的。实际上，混合集落是克隆衍生的，这通过物理分离单细胞来证明。在目前的工作中，没有确定非色素细胞的身份。然而，一种可能的神经命运被提出，因为神经样细胞在培养皿过度生长后发育。即使神经生长因子活性作为培养基的正常成分存在并且在某些情况下作为补充剂添加，也没有形成神经克隆。这些技术允许制备大的，均质的神经嵴细胞群，并提供了一个机会，以检查方面的嵴确定迄今无法研究。
A fundamental question regarding neural crest development is the possible pluripotential nature of this embryonic tissue. As a first step in examining this problem, clonal techniques are used to produce homogeneous populations of crest cells. Primary cultures of these cells are obtained by explanting neural tubes from Japanese quailin vitroand allowing crest cells to migrate away. The explant is removed, the outgrowth is isolated, dissociated with trypsin, and the cells plated at clonal density. Colonies derived in this manner fall into the following categories: all cells of the colony pigmented; none of the cells pigmented; and some of the cells pigmented, the remainder unpigmented. Pigmented colonies generally arise from small, round cells whereas the non-pigmented colonies usually originate from large, flattened polymorphous cells. Differentiation of melanocytes does not preclude their continued proliferation. The pigment phenotype, in addition, is stable through at least 25 generations. That the mixed colonies, in fact, are clonally derived is shown by physically isolating single cells. The identity of the non-pigment cells was not established in the present work. A possible neural fate is suggested, however, since nerve-like cells develop after the petri plates become overgrown. Neural clones did not form even though nerve growth factor activity is present as a normal constituent of the culture medium and was added as a supplement in some instances. These techniques permit the preparation of large, homogeneous populations of neural crest cells and afford an opportunity to examine aspects of crest determination heretofore impossible to study.