Zhang Lanlan, Song Hong, Zhao Xiaojun
Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, 610041 Sichuan, China.
Int J Cell Biol. 2009;2009:747016. doi: 10.1155/2009/747016. Epub 2009 May 27.
Fetal bovine serum (FBS) has been reported to affect chondrocyte biosynthesis in monolayer culture. Insulin-Transferrin-Selenium (ITS) was investigated as a partial replacement for FBS during in vitro culture of rabbit articular chondrocytes in three-dimensional alginate scaffold. Chondrocyte-seeded alginate hydrogels were cultured in Dulbecco's modified Eagle's medium plus 10% FBS, 1% ITS plus 2% FBS, 1% ITS plus 4% FBS, or 1% ITS plus 8% FBS. At designed time point, the Chondrocyte-seeded alginate hydrogels were harvested and evaluated with histological staining, immunohistochemistry, and quantitative gene expression analysis. Viable cell density and cell division were also evaluated. Chondrocytes biosynthesis and cell division in 1% ITS with 2% FBS medium were similar to that in medium added with 10% FBS. For a total culture of 3 weeks, phenotypic gene expression in chondrocyte-seeded hydrogels was maintained at high levels in medium with 1% ITS plus 2% FBS, while it was decreased to varying degrees in the other groups. In conclusion, with 1% ITS, medium with 2% FBS could promote chondrocyte biosynthesis and cell division, and prevented cell dedifferentiation in three-dimensional alginate scaffolds.
据报道，胎牛血清（FBS）会影响单层培养中软骨细胞的生物合成。在三维海藻酸盐支架中对兔关节软骨细胞进行体外培养期间，研究了胰岛素-转铁蛋白-硒（ITS）作为FBS的部分替代物的效果。将接种软骨细胞的海藻酸盐水凝胶分别培养于含10% FBS的杜氏改良 Eagle 培养基、含1% ITS加2% FBS的培养基、含1% ITS加4% FBS的培养基或含1% ITS加8% FBS的培养基中。在设定的时间点，收获接种软骨细胞的海藻酸盐水凝胶，并通过组织学染色、免疫组织化学和定量基因表达分析进行评估。还评估了活细胞密度和细胞分裂情况。在含1% ITS加2% FBS的培养基中，软骨细胞的生物合成和细胞分裂与含10% FBS的培养基中的情况相似。在总共3周的培养过程中，在含1% ITS加2% FBS的培养基中，接种软骨细胞的水凝胶中的表型基因表达维持在高水平，而在其他组中则有不同程度的下降。总之，在含1% ITS时，含2% FBS的培养基可促进软骨细胞的生物合成和细胞分裂，并防止三维海藻酸盐支架中的细胞去分化。
