摘要
目的探讨牛胰岛素-转铁蛋白-硒钠(ITS)对藻酸盐凝胶三维培养条件下兔关节软骨细胞表型的影响。方法分离培养兔关节软骨细胞,将单层培养的P2代细胞接种于藻酸盐凝胶中,分别在DMEM培养液中加入10%胎牛血清(FBS)、0.2%FBS+1%ITS、1%ITS即设为FBS组、ITS/FBS组,ITS组行DNA、蛋白聚糖(GAG)含量测定及组织化学检测。结果软骨细胞在藻酸盐凝胶中始终保持球形样外观,甲苯胺蓝染色及Ⅱ型胶原免疫组化染色呈阳性。3组GAG合成无显著性差异(P>0.05)。培养的第6天ITS组DNA含量低于FBS组(P<0.05),第12天低于ITS/FBS组和FBS组(P<0.05)。结论ITS可促进藻酸盐凝胶中软骨细胞的增殖,并保持其表型。
Objective To investigate the effects of Insulin-Transferrin-Selenium （ITS） on the phenotype of rabbit articular chondro cytes cultured in three-dimensional （3D） alginate hydrogel, Methods Chondrocytes isolated from articular cartilage of rabbit were replica ted in monolayer culture system to obtain the second passage of the cells, which were then suspended in sodium alginate solution for third passage culture. Chondrocyte-seeded alginate hydrogels were maintained in the following culture media： DMEM supplemented respectively with 10% FBS （fetal bovine serum）, DMEM SUDulemented with 1% ITS and 0.2% FBS. or DMEM SuDolemented with 1% ITS.According to the media listed above, the cells were divided into three groups： FBS, ITS/FBS and ITS group. As designed, the cells were harvested from alginate hydrogels at different culture time, and the proliferation and phenotype of the harvested cells were evaluated by de tecting the content of DNA and glycosaminoglycans （GAG）, and also histochemistry staining. Results Chondrocytes could retain their spheroidal morphology when cultured in alginate hydmgels. The cells harvested from hydrogels were positive for toluidine blue and type Ⅱcollagen immunohistochemistry. No significant difference was found in the synthesis of GAG in the three kinds of tested media （ P〉0.05）. The DNA content in samples of ITS group was significantly lower than that of FBS group on day 6 （ P〈0.05）, whereas it was significant ly lower than that of both ITS/FBS group and FBS group on day 12 （P〈0. 05）. Conclusions In alginate hydrogels, ITS can stimulate the proliferation of articular chondrocytes and play a part in maintaining an excellent differentiation phenotype.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2009年第2期159-162,共4页
Medical Journal of Chinese People's Liberation Army
