Zimmermann B, Cristea R
Institut für Anatomie, Freie Universität Berlin, Germany.
Anat Embryol (Berl). 1993 Jan;187(1):67-73. doi: 10.1007/BF00208197.
The effect of dexamethasone on morphogenesis and differentiation of cells obtained from mouse embryos grown at high density in vitro was investigated. Cells from decapitated mouse embryos of day 10 to day 13 were isolated by enzymatic treatment and grown at high density at the medium/air interface in organoid culture. After 28 days in culture, organoid-like structures such as vesicles, gland-like structures and cell aggregates had developed, dependent on the stage of the embryos. After the addition of 10(-7) M dexamethasone, cartilage had formed in cultures of cells from day-10, -11 and -12 embryos. It was maximal in cultures of day-11 cells and rare in day-10 cells. No cartilage was found in cultures of day-13 cells. Cartilage induction was similar in cultures treated with 10(-6) and 10(-7) M dexamethasone, but clearly less in cultures treated with 10(-8) M. Only minute amounts of cartilage were detectable after dexamethasone treatment of cells obtained from decapitated embryos (day-11 and -12) whose limb buds had been cut off. In organoid cultures of pure limb bud cells, 10(-7) M dexamethasone had no influence on chondrogenesis. The results indicate that the cells inducible to form cartilage by dexamethasone originate from the limb buds. Glucocorticoid induction of chondrogenesis has not been described in vivo. The dependency of both chondrogenesis and expression of glucocorticoid receptor on cell density in vitro may be the cause for this effect.
研究了地塞米松对体外高密度培养的小鼠胚胎细胞形态发生和分化的影响。通过酶处理分离出第10至13天断头小鼠胚胎的细胞,并在类器官培养中于培养基/空气界面进行高密度培养。培养28天后,根据胚胎阶段形成了囊泡、腺样结构和细胞聚集体等类器官样结构。添加10^(-7)M地塞米松后,第10、11和12天胚胎细胞培养物中形成了软骨。在第11天细胞培养物中软骨形成最多,在第10天细胞培养物中较少。在第13天细胞培养物中未发现软骨。用10^(-6)M和10^(-7)M地塞米松处理的培养物中软骨诱导情况相似,但用10^(-8)M处理的培养物中明显较少。用地塞米松处理其肢芽已被切除的断头胚胎(第11和12天)获得的细胞后,仅可检测到微量软骨。在纯肢芽细胞的类器官培养中,10^(-7)M地塞米松对软骨形成没有影响。结果表明,可被地塞米松诱导形成软骨的细胞起源于肢芽。体内尚未描述糖皮质激素诱导软骨形成的情况。体外软骨形成和糖皮质激素受体表达对细胞密度的依赖性可能是造成这种效应的原因。