BMP-2体外诱导OPG-/-小鼠BMSCs向成骨方向分化的实验研究

doi:10.3969/j.issn.1674-5817.2012.03.003

实验动物与比较医学 ›› 2012, Vol. 32 ›› Issue (3): 186-190.DOI: 10.3969/j.issn.1674-5817.2012.03.003

BMP-2体外诱导OPG^-/-小鼠BMSCs向成骨方向分化的实验研究

张春¹, 魏琴¹, 王雪梅¹, 匡颖², 段明军¹, 卓菲亚·克依木¹, 姜涛¹, 马嵋¹, 温浩¹

1.新疆医科大学第一附属医院实验动物部, 乌鲁木齐 830054;
2.上海南方模式生物研究中心, 上海 201203

收稿日期:2011-08-29 出版日期:2012-06-25 发布日期:2012-06-25
作者简介:张春(1981-), 男, 兽医硕士, 助理研究员, 主要从事基因工程动物的研究和实验动物管理工作,E-mail: xjyfy_zhang@sina.com
基金资助:
新疆维吾尔自治区科技支疆项目资助(项目编号 201091154)新疆医科大学第一附属医院组织工程专项基金资助(项目编号 2009-ZZGC-04)

Study on BMP-2 on Inducing OPG^-/-BMSCs Differentiate into Osteogenesis in vitro

ZHANG Chun¹, WEI Qin¹, WANG Xue-mei¹, KUANG Ying², DUAN Ming-jun³, ZUOFEIYA·Keyimu¹, JIANG Tao¹, MA Mei¹, WEN Hao¹

1. Department of Experimental Animal, First Teaching Hospital, Xinjiang Medical University, Wulumuqi 830054, China;
2. Shanghai Research Center For Model Organisms, Shanghai 201203, China

Received:2011-08-29 Online:2012-06-25 Published:2012-06-25

摘要/Abstract

摘要： 目的探讨骨形成蛋白-2(BMP-2) 对OPG^-/-小鼠骨髓间充质干细胞(BMSCs)体外定向诱导成骨的能力, 评价BMP-2 活性多肽的成骨诱导性及诱导成骨的剂量依赖性。方法取OPG^-/-小鼠股骨, 分离骨髓基质细胞进行体外培养,传至P₁代时改用条件培养基,在15%FBS培养条件下培养基中分别加入浓度为0.3 μg、0.2 μg、0.1 μg、0.05 μg、0 μg的BMP-2, 并依次记为A、B、C、D 和E 组。细胞培养至1、3、5、7 d, PNPP法测定碱性磷酸酶(ALP)、钙试剂盒测定细胞钙含量、酶联免疫法测定细胞上清中抗酒石酸酸性膦酸酶5b(TRACP5b) 的表达, 检测不同浓度BMP-2 活性多肽体外诱导成骨的能力。结果 ALP活性和钙含量检测显示, A 组和B 组较其他组增加明显 (P<0.05), 但A、B 组间比较差异无统计学意义( P>0.05)。TRACP5b检测显示,A、B、C组第3、5和7 日 TRACP5b表达均低于D、E 组 (P<0.05), 但A和B组差异不显著 (P>0.05)。结论 BMP-2 活性多肽能有效地促进OPG^-/-小鼠BMSCs向成骨方向分化, 其诱导作用存在剂量依赖关系, 最佳诱导剂量为0.2 μg/ml。

关键词: BMP-2, OPG^-/-小鼠, 骨髓间充质干细胞, 诱导成骨

Abstract: Objective To investigate the effect of bone morphogenetic protein-2(BMP-2) on inducing OPG^-/- BMSCs differentiate into the osteogenesis in vitro, and to evaluate the effect of BMP-2 Bioactive Peptides inducing to osteoplast and dose dependent. Methods Got the femur of mice, separated the BMSCs and cultured in vitro. Followed by passaged the BMSCs to P1 using the conditioned medium, and then the BMSCs were added in the BMP-2 consistency of 0.3 μg、0.2 μg、0.1 μg、0.05 μg、0 μg separately cultured with 15% FBS culture medium, marked A, B, C, D, and E group in order. The 3^rd, 5^th, 7^th day after cell culture, the expression of alkaline phosphatase was detected by PNPP method, and calcium content was detected by Calcium kit. The expression of tatrate-resistant acid phosphatase form 5 b in the cell supernatant detected by ELISA, To investigate the effect of different concentration bone morphogenetic protein-2 (BMP-2) on inducing OPG^-/- BMSCs differentiate into the osteogenesis in vitro. Results The activity of alkaline phosphatase and calcium content showed that group A and group B both increase higher than the other group (P<0.05). However no significant differences were detected between the group A and B (P>0.05). TRACP5b displayed, TRACP5b expression in group A, B and C is lower than the other groups in the 3^rd, 5^th, 7^th day (P<0.05), no significant differences were detected between the group A and B (P>0.05). Conclusions The BMP-2 had significant effect on inducing OPG^-/- BMSCs differentiate into the osteogenesis in vitro. The effect dependent on the different concentration of bone morphogenetic protein-2(BMP-2), the optimal concentration is 0.2 μg/ml.

Key words: Bone morphogenetic protein-2(BMP-2), OPG^-/- mouse, Bone marrow derived stroma cell

中图分类号:

Q95-33

张春, 魏琴, 王雪梅, 匡颖, 段明军, 卓菲亚·克依木, 姜涛, 马嵋, 温浩. BMP-2体外诱导OPG^-/-小鼠BMSCs向成骨方向分化的实验研究[J]. 实验动物与比较医学, 2012, 32(3): 186-190.

ZHANG Chun, WEI Qin, WANG Xue-mei, KUANG Ying, DUAN Ming-jun, ZUOFEIYA·Keyimu, JIANG Tao, MA Mei, WEN Hao. Study on BMP-2 on Inducing OPG^-/-BMSCs Differentiate into Osteogenesis in vitro[J]. Laboratory Animal and Comparative Medicine, 2012, 32(3): 186-190.

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BMP-2体外诱导OPG^-/-小鼠BMSCs向成骨方向分化的实验研究

Study on BMP-2 on Inducing OPG^-/-BMSCs Differentiate into Osteogenesis in vitro

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BMP-2体外诱导OPG-/-小鼠BMSCs向成骨方向分化的实验研究

Study on BMP-2 on Inducing OPG-/-BMSCs Differentiate into Osteogenesis in vitro

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BMP-2体外诱导OPG^-/-小鼠BMSCs向成骨方向分化的实验研究

Study on BMP-2 on Inducing OPG^-/-BMSCs Differentiate into Osteogenesis in vitro