碱性成纤维细胞生长因子对体外培养的兔骨髓基质细胞生长特性的影响

目的：研究骨髓基质细胞（BMSC）在体外培养条件下分化为成骨细胞的能力，及碱性成纤维细胞生长因子（bFGF）对其贴附、增殖、分化的影响。方法：将各组BMSC传代后加入bFGF，其浓度分别为0、5、10、50、100、200?ng/ml，用倒置相差显微镜、电镜、四唑盐比色法、碱性磷酸酶（ALP）染色、钙染色和电子探针等方法观察其形态、贴附、生长增殖活性、分化成熟和体外钙化的状况。结果：经10?ng/ml bFGF诱导后，BMSC贴附细胞数量明显增加。当100?ng/ml时，bFGF对BMSC的促增殖作用达到最大值。当加入bFGF时，ALP染色阳性率降低，当bFGF浓度为100?μg/L时阳性率最低（P<0.01）。传代细胞连续培养30?d后钙染色结果呈阳性，电子探针检测矿化区的Ca/P值与羟基磷灰石结晶中Ca/P值相符。电镜观察可见培养的细胞具有典型的成骨细胞形态特征，并能产生胶原纤维。结论：BMSC在体外培养条件下具有分化为成骨细胞的能力，bFGF能够促进细胞的贴壁和增殖，但对其分化成熟具有抑制作用。

Influence of basic fibroblast growth factor in promoting the bone marrow stromal cell growth in vitro

To research the potential ability of bone marrow stromal cells(BMSCs) to differentiate into osteoblasts and the influence of basic fibroblast growth factor(bFGF) on BMSCs adhesion, proliferation and differentiation. Methods：After the BMSCs were cultured successfully, 0, 5, 10, 50, 100, 200?ng/ml bFGF was given respectively. The BMSCs′morphology, adhesion, proliferation and the potentiality of differentiation and calcification in vitro were studied by contrast microscope, electron microscope, MTT test, ALP staining, calcium staining and electronic probe. Results: The adhesion of BMSCs remarkably increased after induced by bFGF at a concentration of 10?ng/ml,　and it reached the peak by bFGF at a concentration of 100?ng/ml. After 100?ng/ml bFGF was added into medium, BMSCs showed a lower positivity rate in ALP staining. After cultured 30 days in conditioned medium, the BMSCs showed apositivity result in Red alizarin calcium staining. The electronic probe result revealed that the main elements of the mineralized nodes were calcium and phosphorus, and this ratio of Ca to P corresponded with the ratio in HA. The cultured BMSCs showed typical osteoblast morphological features in SEM and TEM, in addition, collagen fiber was found in SEM. Conclusion: The BMSCs cultured in vitro can differentiate into osteoblasts. bFGF has a positive influence on BMSCs adhesion and proliferation and a negative influence on their differentiation.