STAGE-RELATED CAPACITY FOR LIMB CHONDROGENESIS IN CELL-CULTURE
STAGE-RELATED CAPACITY FOR LIMB CHONDROGENESIS IN CELL-CULTURE
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DOI：
10.1016/0012-1606(77)90110-5
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发表时间：
1977-01-01
影响因子：
2.7
通讯作者：
REITER, RS
中科院分区：
文献类型：
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作者：
AHRENS, PB;SOLURSH, M;REITER, RS
从不同阶段的鸡胚翅芽中分离细胞，并在培养中生长，以测试它们的软骨分化能力。微团培养物以大于汇合的细胞层开始，其占据培养皿表面的有限区域（10-13 mm 2）。来自阶段24的鸡胚翅芽（在体内出现软骨之前）的细胞在这样的培养物中经历软骨分化。在培养的前1-2天内，细胞形成聚集体（密度比周围非聚集区域大1.5倍的细胞簇）。到第3天，几乎所有的聚集体都分化成软骨结节，这些软骨结节很容易通过它们的阿辛蓝染色（pH 1.0）细胞外基质识别。结节增大，相邻结节开始合并。使用微团培养物来测试来自鸡胚的翅芽细胞的软骨形成能力，所述鸡胚代表肢体发育的不同阶段直至体内软骨出现（阶段17-25）。来自胚胎阶段21-24的细胞形成聚集体，其在体外以相等的能力分化成软骨结节（以结节数/培养物评分）。来自胚胎阶段17-19的细胞以相似的数量形成聚集体，但这些聚集体在常规条件下从未分化成结节。如果暴露于二丁酰环[c]AMP和茶碱，则在第19阶段翅芽细胞培养物中形成的聚集体确实分化成软骨结节。来自第20阶段胚胎的细胞表现出不同的形成软骨结节的能力;显然，这是一个过渡阶段。来自第25阶段胚胎的细胞在体外产生软骨，而不形成聚集体或结节。从胚胎中胚层细胞的软骨分化的模型，提出了，涉及：聚集，收购的能力，以应对环境中的聚集体，细胞内cAMP水平升高，和稳定和表达的软骨表型。
Cells from wing buds of varying-stage chick embryos were dissociated and grown in culture to test their capacity for cartilage differentiation. Micro-mass cultures were initiated with a cell layer greater than confluency, which occupied a restricted area of the culture dish surface (10-13 mm2). Cells from stage 24 chick embryo wing buds (prior to the appearance of cartilage in vivo) undergo cartilage differentiation in such cultures. During the first 1-2 days of culture, cells formed aggregates (clusters of cells with a density 1.5 times greater than that of the surrounding nonaggregate area). By day 3 virtually all aggregates differentiated into cartilage nodules which were easily recognized by their Alcian blue staining (pH 1.0) extracellular matrix. Nodules increased in size and adjacent nodules began to coalesce. Micro-mass cultures were used to test the chondrogenic capacity of wing bud cells from chick embryos representing the different stages of limb development up to the appearance of cartilage in vivo (stages 17-25). Cells from embryo stages 21-24 formed aggregates which differentiated into cartilage nodules in vitro with equal capacity (scored as number of nodules/culture). Cells from embryo stages 17-19 formed aggregates in similar numbers, but these aggregates never differentiated into nodules under routine conditions. Aggregates which formed in cultures of stage 19 wing bud cells did differentiate into cartilage nodules if exposed to dibutyryl cyclic[c]AMP and theophylline. Cells from stage 20 embryos manifested a varying capacity to form cartilage nodules; apparently, this was a transition stage. Cells from stage 25 embryos produced cartilage in vitro without forming either aggregates or nodules. A model for cartilage differentiation from embryonic mesoderm cells is proposed, involving: aggregation, acquisition of the ability to respond to the environment in the aggregate, elevated intracellular cAMP levels, and stabilization and expression of cartilage phenotype.