Ekanayake S, Hall B K
Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada.
Acta Anat (Basel). 1994;151(2):71-9. doi: 10.1159/000147646.
In vitro cloning of embryonic cells is a powerful tool for understanding the mechanism and timing of cell differentiation during development. Although in vitro mass culture of chondrogenic mesenchymal cells and clonal culture of already differentiated chondrocytes have been reported, clonal culture of chondrogenic ectomesenchymal cells at a postmigratory stage but prior to chondrogenic cell condensation has not been reported, nor have cartilage nodules been obtained in clonal culture. We cloned chick mandibular ectomesenchymal cells from HH stage 17 chick embryos. These cells produced clones that underwent chondrogenesis as determined by the presence of alcian blue-staining extracellular matrix and immunohistochemical visualization of type II collagen. Chondrogenic clones formed three-dimensional cartilaginous nodules. This postmigratory mandibular ectomesenchyme is heterogeneous, containing two different types of unipotential cells that give rise to chondrogenic and nonchondrogenic cells, plus bipotential cells that give rise to both chondrogenic and nonchondrogenic cells.
胚胎细胞的体外克隆是理解发育过程中细胞分化机制和时间的有力工具。虽然已有报道称可对软骨生成间充质细胞进行体外大规模培养以及对已分化的软骨细胞进行克隆培养,但尚未见关于迁移后但软骨生成细胞凝聚前的软骨生成外胚间充质细胞克隆培养的报道,也未在克隆培养中获得软骨结节。我们从HH 17期鸡胚中克隆了鸡下颌外胚间充质细胞。这些细胞产生的克隆发生了软骨生成,这通过阿利新蓝染色的细胞外基质的存在以及II型胶原的免疫组织化学可视化得以确定。软骨生成克隆形成了三维软骨结节。这种迁移后的下颌外胚间充质是异质性的,包含两种不同类型的单能细胞,分别产生软骨生成细胞和非软骨生成细胞,以及双能细胞,可同时产生软骨生成细胞和非软骨生成细胞。