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首页> 外文期刊>Cell and Tissue Research >Hypertonic conditions enhance cartilage formation in scaffold-free primary chondrocyte cultures.
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Hypertonic conditions enhance cartilage formation in scaffold-free primary chondrocyte cultures.

机译：高渗条件增强了无支架原代软骨细胞培养物中软骨的形成。

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摘要

The potential of hypertonic conditions at in vivo levels to promote cartilage extracellular matrix accumulation in scaffold-free primary chondrocyte cultures was investigated. Six million bovine primary chondrocytes were cultured in transwell inserts in low glucose (LG), high glucose (HG), or hypertonic high glucose (HHG) DMEM supplemented with fetal bovine serum, antibiotics, and ascorbate under 5 % or 20 % O2 tension with and without transforming growth factor (TGF)-β3 for 6 weeks. Samples were collected for histological staining of proteoglycans (PGs) and type II collagen, analysis by quantitative reverse transcription plus the polymerase chain reaction (RT-PCR) of mRNA expression of aggrecan and procollagen α1 (II) and of Sox9 and procollagen α2 (I), and quantitation of PGs and PG separation in agarose gels. Cartilage tissues produced at 20 % O2 tension were larger than those formed at 5 % O2 tension. Compared with LG, the tissues grew to larger sizes in HG or HHG medium. Histological staining showed the strongest PG and type II collagen staining in cartilage generated in HG or HHG medium at 20 % O2 tension. Quantitative RT-PCR results indicated significantly higher expression of procollagen α1 (II) mRNA in cartilage generated in HHG medium at 20 % O2 tension compared with that in the other samples. TGF-β3 supplements in the culture medium provided no advantage for cartilage formation. Thus, HHG medium used at 20 % O2 tension is the most beneficial combination of the tested culture conditions for scaffold-free cartilage production in vitro and should improve cell culture for research into cartilage repair or tissue engineering.
机译：研究了高渗条件在体内水平上促进无支架原代软骨细胞培养物中软骨细胞外基质积累的潜力。在5％或20％的O2张力下，在补充有胎牛血清，抗生素和抗坏血酸的低葡萄糖（LG），高葡萄糖（HG）或高渗高葡萄糖（HHG）DMEM的Transwell插入物中培养600万牛原软骨细胞。并且在6周内没有转化生长因子（TGF）-β3。收集样品进行蛋白聚糖（PGs）和II型胶原的组织学染色，通过定量逆转录加聚合酶链反应（RT-PCR）进行蛋白聚糖和前胶原α1（II）和Sox9和前胶原α2（I）mRNA表达的分析），以及琼脂糖凝胶中PG的定量和PG分离。 O2张力为20％时产生的软骨组织大于O2张力为5％时形成的软骨组织。与LG相比，组织在HG或HHG培养基中长大。组织学染色显示在20％O2张力下，在HG或HHG培养基中产生的软骨中最强的PG和II型胶原蛋白染色。定量RT-PCR结果表明，与其他样品相比，在20％O2张力下HHG培养基中生成的软骨中前胶原α1（II）mRNA的表达明显更高。培养基中的TGF-β3补充剂对软骨形成没有好处。因此，在20％O2张力下使用的HHG培养基是用于体外无支架软骨生产的测试培养条件的最有益组合，应改善细胞培养以研究软骨修复或组织工程。

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