摘要
背景：研究表明,胰岛素样生长因子Ⅰ和碱性成纤维细胞生长因子对细胞的增殖分化有重要作用,但对人类牙乳头间充质细胞生物学特性有何影响尚不清楚。目的：观察胰岛素样生长因子Ⅰ和碱性成纤维细胞生长因子对体外培养人类牙乳头间充质细胞增殖和分化能力的影响。方法：在建立人类牙乳头间充质细胞培养模型的基础上,①分别用含体积分数为1%或10%胎牛血清的DMEM/F12培养基将两种生长因子配制成不同的实验终浓度,取第4代生长良好的人类牙乳头间充质细胞,分别加入含0,0.1,1,10,100μg/L碱性成纤维细胞生长因子和0,25,50,75,100μg/L胰岛素样生长因子Ⅰ的培养基（0μg/L作对照）,培养96h后四甲基偶氮唑盐法测增殖活性。②设10μg/L碱性成纤维细胞生长因子组、100μg/L胰岛素样生长因子Ⅰ组、碱性成纤维细胞生长因子＋胰岛素样生长因子Ⅰ组和对照组。同上述方法每孔分别加含相应质量浓度生长因子的培养液,分别在培养1,3,5,7d后,四甲基偶氮唑盐法测细胞增殖活性,改良酶动力学法测定细胞碱性磷酸酶活性。结果与结论：在0~100μg/L质量浓度范围时,两种生长因子对人类牙乳头间充质细胞增殖具有促进作用,碱性成纤维细胞生长因子的促增殖作用比胰岛素样生长因子Ⅰ大,联合作用时有协同促增殖作用。碱性成纤维细胞生长因子的最大有效质量浓度为10μg/L,胰岛素样生长因子Ⅰ的最大作用质量浓度为100μg/L。在0~7d时,碱性成纤维细胞生长因子对人类牙乳头间充质细胞的碱性磷酸酶活性影响不明显,随时间的增加,胰岛素样生长因子Ⅰ对细胞碱性磷酸酶活性影响增加,与碱性成纤维细胞生长因子联用时,对增加碱性磷酸酶的活性有协同作用。
BACKGROUND： Previous research has indicated that both insulin-like growth factor Ⅰ （IGF-Ⅰ） and basic fibroblast growth factor （bFGF） play an important role in cell proliferation and differentiation. However, the effect on biological characteristics of human dental papilla mesenchymal cells （hDPMCs） still remains unclear. OBJECTIVE： To research the effect of IGF-Ⅰ and bFGF on the proliferation and differentiation of hDPMCs. METHODS： hDPMCs were isolated and cultured in DMEM/F12 culture media containing 1% or 10% fetal bovine serum. The fourth-passaged hDPMCs were incubated in culture media containing 0, 0.1, 1, 10 and 100 μg/L bFGF and 0, 25, 50, 75 and 100 μg/L IGF-Ⅰ（0 μg/L as control group）, respectively. At 96 hours after culture, proliferative activity was measured with MTT assay. The corresponding growth factor culture media were used in 10 μg/L bFGF group, 100 μg/L IGF-Ⅰgroup, bFGF ＋ IGF-Ⅰ group, and control group, respectively. At days 1, 3, 5, and 7 after culture, the proliferative activity was detected using MTT assay, and alkaline phosphatase （ALP） activity was measured using modified enzyme kinetics method. RESULTS AND CONCLUSION： At the 0-100 μg/L mass concentration scope, both bFGF and IGF-Ⅰcould accelerate proliferation of hDPMCs, and the proliferation ability of bFGF was superior to that of IGF-Ⅰ; moreover, the combination of bFGF and IGF-Ⅰcaused a synergetic action to proliferation of hDPMCs. The maximal valid concentration of bFGF was 10 μg/L, and the maximal action concentration of IGF-Ⅰwas 100 μg/L. At 0-7 days, the effect of bFGF on the ALP activity of hDPMCs was not obvious, but the effect of IGF-Ⅰon ALP activity of hDPMCs became greater with the time passing; furthermore, the combination of bFGF and IGF-Ⅰcould generate a synergetic action on increasing the ALP activity.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第1期70-74,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
教育部高等学校优秀青年教师教学科研奖励计划(2003682)
国家科技部重大基础研究前期专项资金资助项目(2002CCC00700)~~
