Differentiation of muscle, fat, cartilage, and bone from progenitor cells present in a bone-derived clonal cell population: effect of dexamethasone.
Differentiation of muscle, fat, cartilage, and bone from progenitor cells present in a bone-derived clonal cell population: effect of dexamethasone.
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DOI：
10.1083/jcb.106.6.2139
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发表时间：
1988-06
期刊：
影响因子：
--
通讯作者：
Aubin JE
中科院分区：
文献类型：
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作者：
Grigoriadis AE;Heersche JN;Aubin JE
RCJ 3.1是从21天胎鼠颅盖骨分离出的一个克隆衍生细胞群，它表现出与成骨细胞相关的特征，即多边形形态、对甲状旁腺激素的环磷腺苷（cAMP）反应、主要合成I型胶原蛋白以及存在1,25 - 二羟维生素D3调节的碱性磷酸酶活性。当在抗坏血酸、β - 甘油磷酸钠和合成糖皮质激素地塞米松存在的条件下培养时，该克隆以时间依赖的方式分化为四种已知间充质来源的形态不同的表型。早在培养9 - 10天就观察到多核肌细胞，12天后形成含脂脂肪细胞，16天后观察到软骨细胞结节，21天后培养形成矿化骨结节。对分化的细胞类型进行了形态学、组织化学和免疫组织化学鉴定。脂肪细胞和软骨细胞的形成依赖于地塞米松的添加；在没有地塞米松的情况下，肌肉和骨表型也以较低频率表达。性甾体激素孕酮和17β - 雌二醇对该系统的分化没有影响，这表明地塞米松的作用代表了糖皮质激素特有的作用。地塞米松浓度增加（10⁻⁹ - 10⁻⁶ M）会使肌管、脂肪细胞和软骨细胞的数量增加；然而，当连续存在35天时，较低浓度似乎能更好地维持肌肉和脂肪细胞表型。由于骨结节形成的频率太低，未对骨结节进行定量。通过在地塞米松存在的情况下以有限稀释度铺板RCJ 3.1细胞获得的单个细胞，被证明可产生能分化为单一或多种表型的亚克隆。因此，数据表明该克隆细胞系包含间充质祖细胞亚群，它们在糖皮质激素的影响下，可在体外分化为四种不同的细胞类型。因此，它是一种独特的细胞系，将在间充质干细胞分化调控的研究中非常有用。
RCJ 3.1, a clonally derived cell population isolated from 21-d fetal rat calvaria, expresses the osteoblast-associated characteristics of polygonal morphology, a cAMP response to parathyroid hormone, synthesis of predominantly type I collagen, and the presence of 1,25- dihydroxyvitamin D3-regulated alkaline phosphatase activity. When cultured in the presence of ascorbic acid, sodium beta- glycerophosphate, and the synthetic glucocorticoid dexamethasone, this clone differentiated in a time-dependent manner into four morphologically distinct phenotypes of known mesenchymal origin. Multinucleated muscle cells were observed as early as 9-10 d in culture, lipid-containing adipocytes formed after 12 d, chondrocyte nodules were observed after 16 d, and mineralized bone nodules formed after 21 d in culture. The differentiated cell types were characterized morphologically, histochemically, and immunohistochemically. The formation of adipocytes and chondrocytes was dependent upon the addition of dexamethasone; the muscle and bone phenotypes were also expressed at low frequency in the absence of dexamethasone. The sex steroid hormones progesterone and 17 beta-estradiol had no effect on differentiation in this system, suggesting that the effects of dexamethasone represent effects specific for glucocorticosteroids. Increasing concentrations of dexamethasone (10(-9)-10(-6) M) increased the numbers of myotubes, adipocytes, and chondrocytes; however, when present continuously for 35 d, the lower concentrations appeared to better maintain the muscle and adipocyte phenotypes. Bone nodules were not quantitated because the frequency of bone nodule formation was too low. Single cells obtained by plating RCJ 3.1 cells at limiting dilutions in the presence of dexamethasone, were shown to give rise to subclones that could differentiate into either single or multiple phenotypes. Thus, the data suggest that this clonal cell line contains subpopulations of mesenchymal progenitor cells which can, under the influence of glucocorticoid hormones, differentiate in vitro into four distinct cell types. It is, therefore, a unique cell line which will be of great use in the study of the regulation of mesenchymal stem cell differentiation.