Dynamic cultivation with radial flow bioreactor enhances proliferation or differentiation of rat bone marrow cells by fibroblast growth factor or osteogenic differentiation factor

利用径向流生物反应器进行动态培养,可通过成纤维细胞生长因子或成骨分化因子增强大鼠骨髓细胞的增殖或分化。

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Abstract

Dynamic cultivation using a radial flow bioreactor (RFB) has gained increasing interest as a method of achieving bone regeneration. In order to enhance bone generation in large bone defects, it is necessary to use an RFB to expand the primary cells such as bone marrow cells derived from biotissue. The present study aimed to evaluate the cell expansion and osteogenic differentiation of rat bone marrow cells (rBMC) when added to basic fibroblast growth factor containing medium (bFGFM) or osteogenic differentiation factor containing medium (ODM) under dynamic cultivation using an RFB. Cell proliferation was evaluated with a DNA-based cell count method and histological analysis. An alkaline phosphatase (ALP) activity assay and immunohistochemistry staining of osteogenic markers including BMP-2 and osteopontin were used to assess osteogenic differentiation ability. After culture for one week, rBMC cell numbers increased significantly under dynamic cultivation compared with that under static cultivation in all culture media. For different culture media in dynamic cultivation, bFGFM had the highest increase in cell numbers. ALP activity was facilitated by dynamic cultivation with ODM. Furthermore, both BMP-2 and osteopontin were detected in the dynamic cultivation with ODM. These results suggested that bFGFM promotes cell proliferation and ODM promotes osteogenic differentiation of rBMC under dynamic cultivation using an RFB.

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