The effect of recombinant human bone morphogenetic protein-2 on the osteogenic potential of rat mesenchymal stem cells after several passages
The effect of recombinant human bone morphogenetic protein-2 on the osteogenic potential of rat mesenchymal stem cells after several passages
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DOI：
10.1080/17453670710013816
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发表时间：
2007-01-01
影响因子：
3.7
通讯作者：
Ishiguro, Naoki
中科院分区：
文献类型：
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作者：
Ishikawa, Hisato;Kitoh, Hiroshi;Ishiguro, Naoki
由于骨髓间充质干细胞（BMSCs）的成骨潜能随着传代而降低，因此临床应用需要建立一种既能使BMSCs快速扩增又能保持其分化潜能的培养条件。骨形态发生蛋白刺激间充质祖细胞的成骨分化，并增加干细胞数量。因此，我们通过多次传代分析了重组人骨形态发生蛋白-2 （rhBMP-2）对大鼠骨髓间充质干细胞成骨潜能的影响。材料与方法采用体外培养细胞碱性磷酸酶（ALP）活性、上清骨钙素（OC）浓度、ALP和OC mRNA表达水平评价体外成骨分化。在体内成骨方面，将含和不含rhBMP-2的骨髓间充质干细胞通过所有传代植入胸腺小鼠。结果BMP（+）组细胞成骨标志物水平明显高于BMP（-）组，但无论是否添加rhBMP-2，成骨标志物水平均随传代而降低。与体外实验类似，BMP（+）组在所有通道上都有更大程度的骨和软骨组织形成。从我们的结果来看，即使在rhBMP-2存在下传代多次的骨髓间充质干细胞中，成骨潜能也可以保持。这些细胞具有诱导和参与骨形成的能力，可用于临床应用。
Background Since the osteogenic potential of bone marrow derived mesenchymal stems cells (BMSCs) become reduced with passage, establishment of culture condition that permit the rapid expansion of BMSCs while retaining their potential for differentiation is needed for clinical applications. Bone morphogenetic proteins stimulate osteogenic differentiation in mesenchymal progenitor cells as well as increase stem cell numbers. Thus, we analysed the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on the osteogenic potential of rat BMSCs over several passages.Materials and methods Osteogenic differentiation in vitro was evaluated in terms of the alkaline phosphatase (ALP) activity and the osteocalcin (OC) concentration in the supernatants, and the expression of ALP and OC mRNA in the cultured cells. For in-vivo osteogenesis, BMScs cultured with and without rhBMP-2 through all passages were implanted into athymic mice.Results The level of osteogenic markers were significantly higher in the cells of the BMP(+) group than in the cells of the BMP(-) group, although they decreased with passage irrespective of whether or not rhBMP-2 was added. Similar to the in-vitro experiments, there was a greater degree of bone and cartilage tissue formation in the BMP(+) group over all passages.Interpretation From our results, osteogenic potential can be maintained even in BMSCs that have been passaged several times in the presence of rhBMP-2. These cells are capable of inducing and participating in bone formation and can be used for clinical applications.