Francioli Silvia-Elena, Martin Ivan, Sie Christina-Priska, Hagg Rupert, Tommasini Roberto, Candrian Christian, Heberer Michael, Barbero Andrea
Institute for Surgical Research and Hospital Management, University Hospital Basel, Basel, Switzerland.
Tissue Eng. 2007 Jun;13(6):1227-34. doi: 10.1089/ten.2006.0342.
The expansion of chondrocytes in automated bioreactors for clinical use requires that a relevant number of cells be generated, starting from variable initial seeding densities in one passage and using autologous serum. We investigated whether the growth factor combination transforming growth factor beta 1/fibroblast growth factor 2/platelet-derived growth factor BB (TFP), recently shown to enhance the proliferation capacity of human articular chondrocytes (HACs), allows the efficiency of chondrocyte use to be increased at different seeding densities and percentages of human serum (HS). HACs were seeded at 1,000, 5,000, and 10,000 cells/cm2 in medium containing 10% fetal bovine serum or 10,000 cells/cm2 with 1%, 5%, or 10%HS. The chondrogenic capacity of post-expanded HACs was then assessed in pellet cultures. Expansion with TFP allowed a sufficient number of HACs to be obtained in one passage even at the lowest seeding density and HS percentage and variability in cartilage-forming capacity of HACs expanded under the different conditions to be reduced. Instead, larger variations and insufficient yields were found in the absence of TFP. By allowing large numbers of cells to be obtained, starting from a wide range of initial seeding densities and HS percentages, the use of TFP may represent a viable solution for the efficient expansion of HACs and addresses constraints of automated clinical bioreactor systems.
在用于临床的自动化生物反应器中扩增软骨细胞,需要从一个传代中的可变初始接种密度开始并使用自体血清,生成相应数量的细胞。我们研究了最近显示可增强人关节软骨细胞(HACs)增殖能力的生长因子组合转化生长因子β1/成纤维细胞生长因子2/血小板衍生生长因子BB(TFP),是否能在不同接种密度和人血清(HS)百分比下提高软骨细胞的利用效率。将HACs以1000、5000和10000个细胞/cm²接种于含有10%胎牛血清的培养基中,或以10000个细胞/cm²接种于含有1%、5%或10%HS的培养基中。然后在微团培养中评估扩增后HACs的软骨形成能力。使用TFP进行扩增,即使在最低接种密度和HS百分比下,也能在一个传代中获得足够数量的HACs,并减少了在不同条件下扩增的HACs软骨形成能力的变异性。相反,在不使用TFP的情况下,发现变异性更大且产量不足。通过从广泛的初始接种密度和HS百分比开始获得大量细胞,TFP的使用可能是高效扩增HACs的可行解决方案,并解决了自动化临床生物反应器系统的限制。