摘要
目的探讨P17-BMP2对大鼠骨髓间充质干细胞(BMSCs)增殖和成骨分化的影响。方法体外培养Wistar大鼠BMSCs,分4组培养:对照组(A组)、成骨诱导组(B组)、成骨诱导+P17-BMP2组(C组)、单纯P17-BMP2组(D组)。A组:采用普通DMEM培养基培养;B组:含地塞米松10-7mol/L、Vit C 10 mg/L及β-甘油磷酸钠(β-sodium glycerophos-phate,β-GP)10 mmol/L的DMEM培养基培养;C组:成骨诱导液+P17-BMP2(10μg/mL)培养;D组:含P17-BMP2(10μg/mL)的DMEM培养基培养。CCK-8法检测培养第1、3、5、7天时细胞增殖情况;培养1周后,碱性磷酸酶染色(Alkaline phosphatase,ALP),实时定量PCR检测骨钙素(Osteocalcin,OCN)、骨桥蛋白(Osteopontin,OPN)、转录因子(Runt-related transcription factor 2,Runx2)的mRNA表达。结果与A组相比,D组对大鼠BMSCs增殖无影响,而B组对BMSCs细胞增殖具有促进作用,C组细胞增殖能力显著提高;各组碱性磷酸酶染色均为阳性,但染色强度有差异,C组>B组>D组>A组;Q-PCR检测结果显示:与A相比,B组和C组中所有检测基因的表达均显著提高。结论 P17-BMP2能协同成骨诱导剂促进BMSCs的成骨分化。
Objective To investigate the effects of P17-BMP2 peptide on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods BMSCs derived from 4-week-old Wistar rats were isolated and expanded.Four groups were set in this study: Group A,which were cultured in regular medium(DMEM with 10% fetal calf serum);Group B,which were cultured in the regular medium with dexamethasone 107 mol/L,Vitamin C 10 mg/L and beta-glycerophosphate 10 mmol/L(induction medium);Group C,which were cultured in induction medium with P17-BMP2 10 μg/mL;and Group D,which were cultured in regular medium with P17-BMP2 10 μg/mL.Cell proliferation were measured at 1、3、5、7 days after culture by CCK-8 kit.Alkaline phosphatase(AKP) staining were performed 7 days after culture.The expressions of osteocalcin(OCN),osteopontin(OPN),runt-related transcription factor 2(Runx2) were evaluated by Q-PCR.Results Compared with group A,the proliferation of BMSCs were not promoted in group D,but promoted in group B and highly enhanced in group C.The alkaline phosphatase staining were all positive in each group,but the dyeing intensity were different: group C > group B > group D > group A.Compared with group A,the expressions of OCN,OPN and Runx2 were
出处
《组织工程与重建外科杂志》
2012年第2期65-68,共4页
Journal of Tissue Engineering and Reconstructive Surgery
基金
佳木斯大学研究生创新科研项目(YJSCX2011-026JD)
关键词
骨髓间充质干细胞
P-骨形态发生蛋白
增殖
成骨分化
Bone marrow mesenchymal stem cells
P17-BMP2 peptide
Proliferation
Osteogenic differentiation
