细胞高密度培养促进兔关节软骨细胞糖胺多糖合成作用研究

李凯1; 2; 卫小春2; 徐刚2; 邵越峰2; 李鹏翠2; 丁娟2; 杨述华1*

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李凯1,2,卫小春2,徐刚2,邵越峰2,李鹏翠2,丁娟2,杨述华1*.细胞高密度培养促进兔关节软骨细胞糖胺多糖合成作用研究[J].山西医科大学学报,2009,40(4):318~321

细胞高密度培养促进兔关节软骨细胞糖胺多糖合成作用研究

High density culture enhance the glycosaminoglycan synthesis of the in vitro cultured rabbit articular chondrocytes

DOI：

中文关键词: 软骨细胞〓糖胺多糖〓兔〓关节软骨

英文关键词: chondrocytes 〓glycosaminoglycan 〓rabbit 〓articular cartilage

基金项目:国家自然科学基金资助项目（30872616）

作者	单位
李凯1,2,卫小春2,徐刚2,邵越峰2,李鹏翠2,丁娟2,杨述华1*	1华中科技大学同济医学院附属协和医院骨科,武汉430022 2山西医科大学第二临床医学院骨科；*通讯作者,E mail:shuhuayabc@vip.sina.com

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中文摘要:

目的〓研究细胞密度对体外培养兔关节软骨细胞糖胺多糖（glycosaminoglycan，GAG）合成的影响。〓方法〓 1月龄新西兰兔5只，无菌手术切取双膝关节软骨，采用0.4% Pronase酶和0.025%Ⅱ型胶原酶消化分离关节软骨细胞，来源于同一只兔的软骨细胞分为两部分：一部分以密度2×104 /cm2接种，传代时仍以相同密度接种；另一部分在细胞贴壁后人工降低细胞密度至2×103 /cm2培养。原代（P0）和传1代（P1）细胞均在细胞融合后换液，并且于换液后12，24，36，48，60 h分别抽取上清测量 GAG浓度。采用重复测量资料的方差分析，比较低密度培养P0组与高密度培养P0组、P1组上清GAG浓度。〓结果〓低密度培养P0组上清GAG含量显著低于高密度培养P0组（P<0.001），且低于体外培养时间略长的高密度培养P1组软骨细胞（P<0.05）。〓结论〓高密度培养更有利于软骨细胞维持表型，是软骨平面培养的较好方式。

英文摘要:

Objective〓 To study the effect of cell density on glycosaminoglycan(GAG)synthesis of in vitro cultured rabbit articular chondrocytes. 〓Methods〓 Chondrocytes were isolated from the knee joints of 5 one month old rabbits.In high density group,chondrocytes were seeded at 2×104 /cm2 in every passage.In low density group, chondrocyts were seeded at 2×103 /cm2.The morphology of the cells was observed under inverted microscope.After the confluence of cells,the medium was replaced with serum free medium.The supernatants were sampled at 24,36,48,60 h after changing the medium.And the GAG concentrations of supernatants were measured by precipitation with alcian blue. 〓Results〓 The GAG concentration in the supernatant of high density seeded chondrocytes was higher than that of low density seeded chondrocytes in primary culture（P<0.001）.When the high density passage 1 cells reached 100% confluency,chondrocytes experienced longer culture time,but the GAG concentration of the supernatant was higher than that of primary low density seeded chondrocytes（P<0.05）. 〓Conclusion〓 High density culture may enhance the GAG synthesis of in vitro cultured rabbit articular chondrocytes.

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