Differentiation of osteoblasts and in vitro bone formation from murine embryonic stem cells
Differentiation of osteoblasts and in vitro bone formation from murine embryonic stem cells
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DOI：
10.1089/107632700300003323
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发表时间：
2001-02-01
影响因子：
--
通讯作者：
Polak, JM
中科院分区：
文献类型：
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作者：
Buttery, LDK;Bourne, S;Polak, JM
多能胚胎干细胞（ES细胞）具有分化为所有胎儿和成人细胞类型的潜力，可能是组织工程和修复的有用细胞来源。在这里，我们表明，向成骨细胞谱系的ES细胞的分化，可以通过补充含血清的培养基与抗坏血酸，β-甘油磷酸盐，和/或地塞米松/视黄酸或通过与胎鼠成骨细胞共培养增强。ES细胞分化成成骨细胞的特征在于形成离散的矿化骨结节，其由胶原蛋白-1和骨钙素的细胞外基质内的50-100个细胞组成。地塞米松与抗坏血酸和β-甘油磷酸盐的组合诱导的最大数量的骨结节，并依赖于刺激的时间与7倍增加时，加入到ES培养后，但不是之前，14天。与胎儿成骨细胞共培养也提供了一个强有力的刺激成骨分化诱导结节数增加5倍，相对于单独培养的ES细胞。这些数据证明了从多能ES细胞衍生成骨细胞谱系祖细胞的定量测定的应用。这为获得纯化的成骨细胞用于成骨机制的研究和ES细胞在骨组织修复中的应用奠定了基础。
Pluripotent embryonic stem (ES) cells have the potential to differentiate to all fetal and adult cell types and might represent a useful cell source for tissue engineering and repair. Here we show that differentiation of ES cells toward the osteoblast lineage can be enhanced by supplementing serum-containing media with ascorbic acid, beta -glycerophosphate, and/or dexamethasone/retinoic acid or by co-culture with fetal murine osteoblasts. ES cell differentiation into osteoblasts was characterized by the formation of discrete mineralized bone nodules that consisted of 50-100 cells within an extracellular matrix of collagen-1 and osteocalcin. Dexamethasone in combination with ascorbic acid and beta -glycerophosphate induced the greatest number of bone nodules and was dependent on time of stimulation with a seven-fold increase when added to ES cultures after, but not before, 14 days. Co-culture with fetal osteoblasts also provided a potent stimulus for osteogenic differentiation inducing a five-fold increase in nodule number relative to ES cells cultured alone. These data demonstrate the application of a quantitative assay for the derivation of osteoblast lineage progenitors from pluripotent ES cells. This could be applied to obtain purified osteoblasts to analyze mechanisms of osteogenesis and for use of ES cells in skeletal tissue repair.