An ECM substratum allows mouse mesodermal cells isolated from the primitive streak to exhibit motility similar to that inside the embryo and reveals a deficiency in the T/T mutant cells.
An ECM substratum allows mouse mesodermal cells isolated from the primitive streak to exhibit motility similar to that inside the embryo and reveals a deficiency in the T/T mutant cells.
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ECM 培养基允许从原条中分离出的小鼠中胚层细胞表现出与胚胎内部相似的运动性，并揭示了 T/T 突变细胞的缺陷。
DOI：
10.1242/dev.100.4.587
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发表时间：
1987
期刊：
影响因子：
4.6
通讯作者：
N. Nakatsuji
中科院分区：
文献类型：
--
作者：
K. Hashimoto;H. Fujimoto;N. Nakatsuji
中胚层细胞层是由妊娠第7天小鼠胚胎原条区的细胞侵入和迁移而形成的。为了研究中胚层细胞在发育过程中的迁移机制，将从原条中分离的中胚层细胞培养在不同的基质上，并用延时录像系统分析细胞的行为和运动。细胞外基质（ECM）包被的培养皿（牛角膜内皮细胞产生的ECM）表面上的中胚层细胞显示出广泛的迁移，平均迁移率约为100%。50微米h-1。它们还表现出频繁的细胞分裂，并表现出板状伪足的接触麻痹和运动的接触抑制。然而，在塑料或玻璃表面上，中胚层细胞变得更扁平，运动性更差（约为100%）。20-30微米h-1）。ECM上的细胞形状和平均运动速率与原位非常相似，如先前研究中所研究的（Nakatsuji，Snow & Wylie，1986）。因此，这种培养条件可以为分析小鼠胚胎正常和异常形态发生运动的细胞基础提供一个有用的实验系统。采用这样的培养系统，我们研究了来自Brachyury（T）突变纯合子胚胎的中胚层细胞的运动性，该突变在子宫内的妊娠中期是致命的。组织学观察表明，T/T胚胎的异常形态发生可能是由原始条纹中胚层细胞迁移缺陷引起的。当将来自第8-9天的T/T突变胚胎的原条的中胚层细胞在ECM基质上培养时，与来自正常胚胎的细胞相比，细胞迁移的平均速率显著降低。结果支持延迟迁移的突变中胚层细胞的一个重要因素，造成异常的形态发生的想法。
The mesodermal cell layer is created by ingression and migration of the cells from the primitive streak region in mouse embryos on day 7 of pregnancy. In order to study the mechanisms of mesodermal cell migration during development, the mesodermal cells isolated from the primitive streak were cultured on various substrata, and cell behaviour and motility were analysed with a time-lapse video system. The mesodermal cells on the surface of extracellular matrix (ECM)-coated dishes (ECM produced by bovine corneal endothelial cells) showed extensive migration at a mean rate of approx. 50 micron h-1. They also showed frequent cell division and exhibited contact paralysis of lamellipodia and contact inhibition of movement. On plastic or glass surfaces, however, the mesodermal cells became more flattened and less motile (approx. 20-30 micron h-1). Cell shape and mean rate of movement on the ECM were very similar to those in situ, as investigated in a previous study (Nakatsuji, Snow & Wylie, 1986). Therefore, this culture condition could provide a useful experimental system for analysing the cellular basis of normal and abnormal morphogenetic movements in mouse embryos. Employing such a culture system, we studied motility of the mesodermal cells from embryos homozygous for Brachyury (T) mutation, which are lethal at the midgestation stage in utero. Histological observations have suggested that anomalous morphogenesis of the T/T embryos may be brought about by defects in migration of the mesodermal cells derived from the primitive streak. When mesodermal cells from the primitive streak of the T/T mutant embryos on days 8-9 were cultured on the ECM substratum, mean rate of cell migration was significantly reduced compared to cells from normal embryos. Results support the idea of retarded migration by the mutant mesodermal cells as an important factor causing abnormalities in morphogenesis.