FBS对成骨生长肽促进BMSCs增殖分化的影响

发布时间：2018-04-13 01:41

本文选题：FBS + 成骨生长肽　；参考：《中国修复重建外科杂志》2015年02期

【摘要】：目的探讨培养液中不同浓度FBS对成骨生长肽(osteogenic growth peptide,OGP)促进BMSCs增殖和分化的影响。方法取8只5周龄SD大鼠四肢骨,贴壁法分离纯化BMSCs并传代,倒置相差显微镜下观察细胞形态。取第3代BMSCs分组培养:分别采用浓度为1×10-10、1×10-9和1×10-8 mol/L的OGP进行培养,以正常培养细胞作为对照组;同时每组设FBS浓度为0、2%、5%、8%和10%5个梯度。培养后1、3、5、7、9、12 d采用MTT法检测细胞增殖,9 d时采用对硝基苯磷酸二钠法测定早期分化指标细胞内ALP活性。结果倒置相差显微镜下观察BMSCs贴壁生长,增殖迅速,呈纤维状涡旋生长,形态典型。MTT检测示,FBS低于5%时各组细胞不能持续增殖,当FBS浓度为8%以上时细胞可持续增殖;OGP 1×10-8 mol/L和1×10-9 mol/L组在FBS各浓度中促增殖效果均大于对照组(P0.05);FBS浓度低于10%时,OGP 1×10-8 mol/L组促增殖效果显著优于其余OGP浓度组(P0.05),但FBS浓度为10%时OGP 1×10-8 mol/L组无促增殖优势。ALP检测示,各组组内随FBS浓度增加,ALP活性增加(P0.05);当FBS浓度为5%、8%时,各OGP浓度组ALP活性均大于对照组(P0.05),且OGP 1×10-8 mol/L组最高(P0.05);当FBS浓度为10%时,各OGP组ALP活性仍大于对照组(P0.05),但OGP1×10-8 mol/L组与OGP 1×10-9 mol/L组差异无统计学意义(P0.05)。结论 8%FBS浓度为OGP促进BMSCs增殖分化的最佳血清浓度,且OGP促进BMSCs增殖分化的最适浓度为1×10-8 mol/L。
[Abstract]:Objective to investigate the effects of different concentrations of FBS in culture medium on the proliferation and differentiation of osteogenic growth peptidine (OGP) induced by osteogenic growth peptide (OGP).Methods the limb bones of 8 5-week-old SD rats were isolated and purified by adherent method. The morphology of the cells was observed by inverted phase contrast microscope.The third passage of BMSCs was divided into three groups: 1 脳 10 ~ (-10) (1 脳 10 ~ (-9)) and 1 脳 10 ~ (-8) mol/L OGP, respectively, and normal cultured cells were used as the control group, and each group was divided into two groups: the FBS concentration was 0 脳 10 ~ (-10) (1 脳 10 ~ (-9)) and 10 ~ (-8) mol/L, respectively.MTT assay was used to detect cell proliferation at 12 days after culture. The ALP activity was measured by p-nitrobenzene disodium phosphate assay at 9 days after culture.Results the adherent growth of BMSCs was observed under inverted phase contrast microscope, and the proliferation was rapid and fibrous vortex was observed.When the concentration of FBS was more than 8%, the proliferation effect of OGP1 脳 10-8 mol/L and 1 脳 10-9 mol/L group was higher than that of the control group P0.05FBS concentration lower than that of the control group when the concentration of OGP1 脳 10-8 mol/L was lower than that of the other OGP group, but FBS concentration was higher than that of the other OGP group.OGP 1 脳 10 ~ (-8) mol/L group had no proliferative advantage.When the concentration of FBS was 5, the activity of ALP in each OGP group was higher than that in the control group, and the highest level of P0.05in the OGP 1 脳 10-8 mol/L group, when the concentration of FBS was 10, the highest level of P0.05 was found in the group with OGP 1 脳 10 ~ (-8) mol/L, when the concentration of FBS was 10 ~ (-5), the activity of ALP in each group was higher than that in the control group.The activity of ALP in the OGP group was still higher than that in the control group (P0.05), but there was no significant difference between the OGP1 脳 10-8 mol/L group and the OGP 1 脳 10-9 mol/L group.Conclusion 8s concentration is the best serum concentration for OGP to promote the proliferation and differentiation of BMSCs, and the optimal concentration of OGP for BMSCs proliferation and differentiation is 1 脳 10 ~ (-8) mol 路L ~ (-1) 路L ~ (-1) 路L ~ (-1).
【作者单位】：山东中医药大学;山东中医药大学附属医院骨科;山东省医科院基础医学研究所;
【基金】：山东省中医药科技发展计划资助项目(2013-219) 济南市科技支撑计划资助项目(201102016) 山东省自然科学基金资助项目(ZR2014HM085)~~
【分类号】：R329.2

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