Sextier-Sainte-Claire Deville F, Ziller C, Le Douarin N
Institut d'Embryologie cellulaire et moléculaire du CNRS, Nogent-sur-Marne, France.
Brain Res Dev Brain Res. 1992 Mar 20;66(1):1-10. doi: 10.1016/0165-3806(92)90134-i.
The developmental potentialities of single truncal neural crest derived cells were analysed in clonal cultures. The clone-forming ability and differentiation potential of crest cells migrating through the somitic mesoderm of 3-day-old embryos (E3) and of non-neuronal cells of dorsal root ganglia taken at E6-14 were compared. Since most of the cells present in the sclerotomal and rostral parts of the somite at E3 become later on incorporated into the spinal ganglia, one can consider that these two cell populations represent the same derivatives of the trunk neural crest at different developmental stages. After 10 days in vitro, the size of clones and their phenotypic composition varied noticeably, revealing a certain heterogeneity in the founder cell populations in terms of developmental potencies. Clones obtained from migrating neural crest cells at E3 were often large (greater than 1000 cells) and many of them contained neuronal and non-neuronal cells. Dorsal root ganglion cells produced mostly small clones (less than 100 cells) in which only non-neuronal (i.e. glial) phenotypes were expressed. Therefore, both the capacity for proliferation and the differentiation ability of cloned neural crest derived cells decrease considerably with increasing embryonic age. This is even more striking if these results are compared with those obtained previously in our laboratory with single cells cultures of E2 cephalic neural crest. In the latter case, both clone sizes and cellular diversity within the colonies were much higher than with E3 truncal crest and dorsal root ganglia (DRG) non-neuronal cells. The second result of the present work concerns the differentiation of the dormant autonomic neuronal precursors of the DRG. It has been established previously that the non-neuronal cells of the DRG include adrenergic precursors than can differentiate in mass culture of dissociated DRG cells. We show that these cells never differentiate in clonal cultures but depend upon the cell density of the culture. This suggests that cell to cell interaction between crest derived cells are critical in eliciting the differentiation of the adrenergic phenotype.
在克隆培养中分析了单个躯干神经嵴衍生细胞的发育潜能。比较了迁移通过3日龄胚胎（E3）体节中胚层的嵴细胞以及在E6 - 14获取的背根神经节非神经元细胞的克隆形成能力和分化潜能。由于E3时存在于体节硬骨部和头侧部分的大多数细胞后来会并入脊髓神经节，因此可以认为这两个细胞群体代表了躯干神经嵴在不同发育阶段的相同衍生物。体外培养10天后，克隆的大小及其表型组成有明显差异，这表明起始细胞群体在发育潜能方面存在一定的异质性。从E3迁移的神经嵴细胞获得的克隆通常较大（超过1000个细胞），其中许多包含神经元和非神经元细胞。背根神经节细胞大多产生小克隆（少于100个细胞），其中仅表达非神经元（即神经胶质）表型。因此，随着胚胎年龄的增加克隆神经嵴衍生细胞的增殖能力和分化能力均显著下降。如果将这些结果与我们实验室之前用E2头侧神经嵴单细胞培养获得的结果进行比较，这种情况就更加明显。在后一种情况下，克隆大小和集落内的细胞多样性均远高于E3躯干嵴和背根神经节（DRG）非神经元细胞。本研究的第二个结果涉及DRG中休眠自主神经元前体的分化。先前已经确定，DRG的非神经元细胞包括肾上腺素能前体，其可在解离的DRG细胞的大量培养中分化。我们表明，这些细胞在克隆培养中从不分化，但取决于培养物的细胞密度。这表明嵴衍生细胞之间的细胞间相互作用对于引发肾上腺素能表型的分化至关重要。
