CAJ | 학술논문

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0,0.1,1,10,100 μg/L碱性成纤维细胞生长因子和0,25,50,75,100 μg/L胰岛素样生长因子Ⅰ的培养基(0 μg/L作对照),培养96 h后四甲基偶氮唑盐法测增殖活性.②设10 μg/L碱性成纤维细胞生长因子组、100 μg/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1,3,5,7 d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0～100 μg/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10 μg/L,胰岛素样生长因子Ⅰ的最大作用质量浓度为100 μg/L.在0～7 d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.
배경:연구표명,이도소양생장인자Ⅰ화감성성섬유세포생장인자대세포적증식분화유중요작용,단대인류아유두간충질세포생물학특성유하영향상불청초.목적:관찰이도소양생장인자Ⅰ화감성성섬유세포생장인자대체외배양인류아유두간충질세포증식화분화능력적영향.방법:재건립인류아유두간충질세포배양모형적기출상,①분별용함체적분수위1%혹10%태우혈청적DMEM/F12배양기장량충생장인자배제성불동적실험종농도,취제4대생장량호적인류아유두간충질세포,분별가입함0,0.1,1,10,100 μg/L감성성섬유세포생장인자화0,25,50,75,100 μg/L이도소양생장인자Ⅰ적배양기(0 μg/L작대조),배양96 h후사갑기우담서염법측증식활성.②설10 μg/L감성성섬유세포생장인자조、100 μg/L이도소양생장인자Ⅰ조、감성성섬유세포생장인자+이도소양생장인자Ⅰ조화대조조.동상술방법매공분별가함상응질량농도생장인자적배양액,분별재배양1,3,5,7 d후,사갑기우담서염법측세포증식활성,개량매동역학법측정세포감성린산매활성.결과여결론:재0～100 μg/L질량농도범위시,량충생장인자대인류아유두간충질세포증식구유촉진작용,감성성섬유세포생장인자적촉증식작용비이도소양생장인자Ⅰ대,연합작용시유협동촉증식작용.감성성섬유세포생장인자적최대유효질량농도위10 μg/L,이도소양생장인자Ⅰ적최대작용질량농도위100 μg/L.재0～7 d시,감성성섬유세포생장인자대인류아유두간충질세포적감성린산매활성영향불명현,수시간적증가,이도소양생장인자Ⅰ대세포감성린산매활성영향증가,여감성성섬유세포생장인자련용시,대증가감성린산매적활성유협동작용.
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.