C2C12细胞体外微型灌注生物反应器培养初探

Study of C2C12 cells perfusion micro-bioreactor culture in vitro

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。

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 （TCC2C12, 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% mediareplaced 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.