C2C12细胞体外微型灌注生物反应器培养初探

目的 研究组织工程种子细胞C2C12在微型灌注生物反应器中培养的特点,以期探索一种有效的种子细胞体外培养方法.方法 在自行研制的微型灌注生物反应器中以20%换液量/天、30%换液量/天两种条件进行C2C12细胞培养,通过细胞形态、生长曲线、细胞群体倍增时间及生化代谢离线检测等对培养情况进行研究.结果 微型灌注生物反应器培养较传统的细胞培养瓶培养,培养基使用效率提高;在细胞形态无显著性变化的情况下,细胞群体倍增时间显著缩短.而且,C2C12细胞的生化代谢在20%换液量/天灌注条件下较30%换液量/天的为佳.结论 C2C12细胞的灌注生物反应器培养方式较传统的细胞培养瓶培养方式优越,同时,20%换液量/天的灌注条件较30%换液量/天为佳.该灌注培养体系值得进一步深入研究.

Study of C2C12 cells perfusion micro-bioreactor culture in vitro

Objective To investigated the in vitro culture of C2C12 cells,seed cells for bone tissue engineering,in a self-manufactured perfusion micro-bioreactor in vitro.Methods The flow rates were set at 20% media-replaced per day and 30% media-replaced per day.The effects were showed by cellular morphology,culture curve,doubling time and off-line metabolism analysis.Results In perfusion culture groups(BR-C2C12-20 and BR-C2C12-30),the media consumptions were less than conventional static culture group(TC-C2C12,as control).And,the doubling times in perfusion groups were significant shorter than that in control group,while the cellular morphologies were not changed obviously.Within the perfusion groups,the doubling time of BR-C2C12-30 was less than BR-C2C12-20,and the metabolism conditions of BR-C2C12-20 were better than BR-C2C12-30.Conclusion As far as C2C12 cells were concerned,the perfusion micro-bioreactor culture was more efficient than conventional static culture.Also,the culture condition of 20% media-replaced per day was better than that of 30% media-replaced per day for C2C12 cells.This culture system was worthy to be studied further in the future.