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两种不同培养方法对破骨细胞ATPase a3基因表达和骨吸收活性的影响
引用本文:杜文喜,肖鲁伟,吴承亮,厉驹,童培建. 两种不同培养方法对破骨细胞ATPase a3基因表达和骨吸收活性的影响[J]. 浙江中医药大学学报, 2009, 33(1): 28-32
作者姓名:杜文喜  肖鲁伟  吴承亮  厉驹  童培建
作者单位:浙江中医药大学附属第一医院,杭州,310006
摘    要:[目的]探讨不同方法培养下的大鼠破骨细胞(osteoclast,OC)骨吸收功能的差异,以及骨吸收关键基因ATPasea3的mR—NA表达水平的差异,为体外实验奠定基础。[方法]机械分离和诱导培养,即从新生24h的大鼠长干骨骨髓腔内壁机械分离成熟OC和1,25(OH)2D3诱导大鼠骨髓细胞形成破骨样细胞(osteoclast like cell,OLC),对获得的OC进行形态和骨吸收功能观察,并测定OC骨吸收关键基因ATPasea3的mRNA表达水平的差异。[结果]OC和OLC均为抗酒石酸酸性磷酸酶(tartrate resistant acid phosphatase,TRAP)染色阳性的多核巨细胞,与细胞共培养骨片上可形成骨吸收陷窝;诱导法培养出的OLC数目多于机械分离法,但诱导早期陷窝较之小而浅,诱导后期基本接近OC;OC骨吸收关键基因ATPase a3的mRNA,在机械分离8h与诱导培养6天的细胞表达量无显著差异,但都远少于培养8天的表达量。[结论]诱导法可以培育出大量的OLC,优于机械分离法,但早期骨吸收功能较弱,OC骨吸收功能与其核数相关。后期的OCL与机械分离OC接近,可以用于各类实验。

关 键 词:破骨细胞  破骨样细胞  骨吸收  ATPase  a3

Effect of Two Different Cultural Methods on Osteoclast Bone Absorption Activity and Expression of ATPase a3 Gene
Du Wenxi,Xiao Luwei,Wu Chengliang,et alThe First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou. Effect of Two Different Cultural Methods on Osteoclast Bone Absorption Activity and Expression of ATPase a3 Gene[J]. Journal of Zhejiang University of Traditional Chinese Medicine, 2009, 33(1): 28-32
Authors:Du Wenxi  Xiao Luwei  Wu Chengliang  et alThe First Affiliated Hospital of Zhejiang Chinese Medical University  Hangzhou
Abstract:[Objective] To investigate the difference of bone Absorption function of the rat osteoclast(OC) in different cultural methods, and the difference in the levels of mRNA expression of bone Absorption key gene, ATPase a3 gene, and provide the basis for further investigation on the OC in vitro. [Methods] with mechanical separation method, mature osteoclast was separated from inner wall of 24 hours newborn rat long bones medullary cavity; and induction culture methods, bone marrow cell was induced to osteoclast ...
Keywords:ATPasea3
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