Enhancement of osteoblastic differentiation of mesenchymal stromal cells cultured by selective combination of bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor-2 (FGF-2)
Enhancement of osteoblastic differentiation of mesenchymal stromal cells cultured by selective combination of bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor-2 (FGF-2)
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DOI：
10.1002/term.41
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发表时间：
2007-07-01
影响因子：
3.3
通讯作者：
Ohgushi, Hajime
中科院分区：
文献类型：
--
作者：
Maegawa, Naoki;Kawamura, Kenji;Ohgushi, Hajime
众所周知，骨髓中含有间充质基质细胞（MSC），当在含有抗坏血酸、β-甘油磷酸盐和地塞米松的成骨培养基中培养时，MSC可显示成骨分化。这种分化导致成骨细胞的出现，以及骨基质的形成;因此，可以通过MSC培养来制造体外培养的骨（成骨细胞/骨基质）。这种类型的培养骨已经用于涉及骨科问题的临床病例。为了提高培养骨的治疗效果，我们研究了有助于广泛成骨细胞分化的培养条件。原代培养大鼠骨髓以扩增MSCs数量，并在成骨培养基中进一步培养12天。还在补充有骨形态发生蛋白-2（BMP-2）或成纤维细胞生长因子（FGF-2）或两种补充物的顺序组合的培养基中进行培养。其中，连续补充FGF-2和BMP-2显示出高碱性磷酸酶活性、足够的骨特异性骨钙素表达和MSC培养物的丰富的骨基质形成。这些数据表明，在培养的早期阶段，通过补充FGF-2，响应细胞或未成熟成骨细胞的数量增加，并且这些细胞可以显示成骨细胞分化，在后期阶段，BMP-2增强了这些细胞的分化能力。这些细胞因子的顺序补充到MSC培养可能是适合于理想的培养骨用于骨组织工程。版权所有（C）2007约翰威利父子有限公司
It is well known that bone marrow contains mesenchymal stromal cells (MSCs), which can show osteoblastic differentiation when cultured in osteogenic medium containing ascorbic acid, beta-glycerophosphate and dexamethasone. The differentiation results in the appearance of osteoblasts, together with the formation-of bone matrix; thus, in vitro cultured bone (osteoblasts/bone matrix) could be fabricated by MSC culture. This type of cultured bone has already been used in clinical cases involving orthopaedic problems. To improve the therapeutic effect of the cultured bone, we investigated the culture conditions that contributed to extensive osteoblastic differentiation. Rat bone marrow was primarily cultured to expand the number of MSCs and further cultured in osteogenic medium for 12 days. The culture was also conducted in a medium supplemented with either bone morphogenetic protein-2 (BMP-2) or fibroblast growth factor (FGF-2), or with sequential combinations of both supplements. Among them, the sequential supplementation of FGF-2 followed by BMP-2 showed high alkaline phosphatase activity, sufficient bone-specific osteocalcein expression and abundant bone matrix formation of the MSC culture. These data implied that the number of responding cells or immature osteoblasts was increased by the supplementation of FGF-2 in the early phase of the culture and that these cells can show osteoblastic differentiation, of which capability was-augmented by BMP-2 in the late phase. The sequential supplementation of these cytokines into MSC culture might be suitable for the fabrication of ideal cultured bone for use in bone tissue engineering. Copyright (C) 2007 John Wiley & Sons, Ltd.