体外培养的大鼠骨髓基质细胞的生物学特性

Biological characteristics of stromal cells derived from rat bone marrow in vitro

目的 :研究体外培养的大鼠骨髓基质细胞的生物学特性及其成骨细胞表型特征。方法 :分离大鼠四肢长骨骨髓基质细胞 ,常规和矿化培养液培养 ,测定细胞生长曲线 ,碱性成纤维细胞生长因子 (bFGF)、重组人骨形成蛋白 -2 (rhBMP -2 )及矿化培养液对骨髓基质细胞碱性磷酸酶 (ALP)活性的影响 ,观察矿化结节的形成。结果 :大鼠骨髓基质细胞群体倍增时间 (DT )为 66.1h ;在矿化液培养条件下细胞DT为 5 0 .3h。bFGF和rhBMP -2均不同程度增加骨髓基质细胞碱性磷酸酶的活性。矿化液有较明显的增加骨髓基质细胞碱性磷酸酶活性的作用 ,用茜素红染色法显示较多的深红色矿化结节 ;常规培养条件下大鼠骨髓基质细胞生长虽呈现密集单层 ,没有矿化结节形成。结论 :大鼠骨髓基质细胞在体外可以稳定传代培养 ,在矿化培养液诱导培养条件下可以向成骨细胞分化 ,bFGF和rhBMP -2有助于骨髓基质细胞表现成骨细胞表型特征 ,提示骨髓基质细胞在骨移植研究领域有重要的应用价值。

砄bjective:To study the biological characteristics and mineralization activity of strom cells (MSC) isolated from rat bone marrow.Methods: MSCs were isolated from rat bone marrow using gradient centrifuge (460 g,20 min) with 70% percoll and cultured with DMEM. The cells were then incubated with mineralization culture medium (MCM) in the presence of Dex (10～7 mol/L), β GP (10 mmol/L) and AcA (50 g/L).Cell growth was studied with cell counting,alkaline phosphatase (ALP) expression with a kit and mineralization with a saturated solution of alizarin red S (pH 4.3) stainning.Results: The isolated cells from bone marrow of rat were epithelial like. The doubling time (DT) of the cells in MCM or DMEM were 50.3 h and 66.1 h respectively. Mineralization was observed after the cells had been cultured in MCM for 28 days. bFGF and rhBMP 2 increased ALP expression of MSC in a dosage dependent manner. Conclusion: In vitro cultured MSCs may be induced to differentiate towards osteoblasts.