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目的探讨不同浓度血清、不同体外培养时间对L929系小鼠成纤维细胞表达产物Ⅰ、Ⅲ型胶原及其比值的影响。方法采用双抗体夹心法检测细胞培养液中Ⅰ、Ⅲ型胶原的含量,并同时计算出Ⅰ/Ⅲ型胶原的比值。结果除0%血清浓度组的Ⅰ、Ⅲ型胶原呈现无明显变化的微弱升高趋势外,其余各血清浓度组的Ⅰ、Ⅲ型胶原均处于增加状态,且在6~8 d时增加最为明显;另外,Ⅰ/Ⅲ型胶原比值随着血清浓度和细胞生长时间不同,表现出不同的变化趋势。其中20%和40%血清浓度组随着培养细胞生长时间延长,Ⅰ/Ⅲ型胶原比值逐渐增加;而其他血清浓度组,随着细胞培养时间的延长,Ⅰ/Ⅲ型胶原比值增加并不明显。结论 L929细胞在不同体外培养时间和不同浓度血清下其表达产物Ⅰ、Ⅲ型胶原含量及其比值有一定变化,为今后优化体外细胞培养条件提供了相关的实验数据。
Objective To investigate the effect of different concentrations of serum and culture in vitro on the expression of collagen type Ⅰ and Ⅲ and the ratio of fibroblasts in L929 mice fibroblasts. Methods The content of type I and type III collagen in cell culture medium was detected by double antibody sandwich method, and the ratio of type Ⅰ / Ⅲ collagen was also calculated. Results In addition to the 0% serum concentration of type Ⅰ, Ⅲ collagen showed no significant change in the weak upward trend, the remaining serum concentrations of type Ⅰ, type Ⅲ collagen are in an increase state, and 6 to 8 d increased most significantly In addition, the type Ⅰ / Ⅲ collagen ratio with the serum concentration and cell growth time showed different trends. In 20% and 40% of the serum concentration group, the ratio of type Ⅰ / Ⅲ collagen gradually increased with the growth of cultured cells, while in other serum concentration groups, the ratio of type Ⅰ / Ⅲ collagen increased with the increase of cell culture time . CONCLUSION: L929 cells have some changes in the expression of collagen type I and type III and their ratios in different culture time and different concentrations of serum, and provide relevant experimental data for the future optimization of cell culture conditions in vitro.