使用旋转式生物反应器体外扩增人表皮细胞

Amplification of Human Epidermal Cells by a Rotary Bioreactor in Vitro

目的:使用Cytodex-3微载体和高截面纵横比的旋转式生物反应器容器作为培养系统大规模扩增人表皮细胞(hECs)。方法:使用中性蛋白酶和胰蛋白酶-EDTA两步骤法从人皮肤中分离出人表皮细胞,使用DIL标记细胞后结合微载体后在旋转式生物反应器(RCCS)中培养,细胞贴附微载体的生长状态使用倒置显微镜,扫描电镜观测。并且分析细胞群体倍增时间来比较微重力培养与平面培养的体外增殖能力差异。结果:在旋转式生物反应器的微重力培养体系中,人表皮细胞能快速贴附到微载体表面,在培养过程中达到很大的细胞密度,并且表现出很强的增殖能力和细胞活性。结论:使用旋转式生物反应器和微载体悬浮培养人表皮细胞,是大量制备皮肤组织工程种子细胞的一种有效方法。

Objective： We attempted to produce human epidermal cells （hECs） on a large scale by Rotary Cell Culture System （RCCS）. In this cultural system we used Cytodex-3 microbeads as microcarriers and high aspect ratio vessel RCCS as cultural system. Methods：The hECs were obtained from human skin by using the method of digesting human foreskin with Dispase II and Trypsin-EDT and cultured in vitro and labeled with DIL, then combined with Cytodex-3 in RCCS. The characteristics of hECs combined with microcarriers were observed with Inverted microscope, and scanning electronic microscope （SEM）. The proliferative capacities of microgravity cultivation and routine cultivation were compared by group doubling time. Results：Our results showed that hECs rapidly attached to the microcarriers in the RCCS, achieving high cell densities, hECs cultured on the microbeads in the RCCS remained actively proliferating potentials. Conclusion： Using biological reactor and method for preparation a large number of seed cells for tissue engineering.