Ayyildiz-Tamis Duygu, Avcı Kamuran, Deliloglu-Gurhan S Ismet
Engineering Faculty, Department of Bioengineering, Ege University, Izmir, 35100, Bornova, Turkey,
In Vitro Cell Dev Biol Anim. 2014 Mar;50(3):221-31. doi: 10.1007/s11626-013-9717-y. Epub 2013 Dec 20.
Microcarriers provide large adhesion area allowing high cell densities in bioreactor systems. This study focused on the investigation of cell adhesion and cell growth characteristics of both anchorage-dependent CHO-K1 and anchorage-independent Ag8 myeloma cell lines cultivated on four different microcarriers (Biosilon®, Microhex®, Cytodex 3®, Cytoline 2®) by considering the cell kinetics and physiological data. Experiments were performed in both static and agitated cell culture systems by using 24-well tissue culture plates and then 50-ml spinner flasks. In agitated cultures, the highest specific growth rates (0.026 h for CHO-K1 and 0.061 h for Ag8 cell line) were obtained with Cytodex 3® and Cytoline 2® microcarriers for CHO-K1 and Ag8 cell line, respectively. Metabolic characteristics showed some variation among the cultures with the four microcarriers. The most significant being the higher production of lactate with microcarriers with CHO-K1 cells relative to the Ag8 cells. SEM analyses revealed the differences in the morphology of the cells along with microcarriers. On Cytodex 3® and Cytoline 2®, CHO-K1 cells attached to the substratum through long, slender filopodia, whereas the cells showed a flat morphology by covering the substratum on the Biosilon® and Microhex®. Ag8 cells maintained their spherical shapes throughout the culture for all types of microcarriers. In an attempt to scale-up, productions were carried out in 50-ml spinner flasks. Cytodex 3® (for CHO-K1 cells) and Cytoline 2® (for Ag8 cells) were evaluated. The results demonstrate that high yield of biomass could be achieved through the immobilization of the cells in each culture system. And cell cultures on microcarriers, especially on Cytodex 3® and Cytoline 2®, represented a good potential as microcarriers for larger scale cultures of CHO-K1 and Ag8, respectively. Moreover, owing to the fact that the cell lines and culture media are specific, outcomes will be applicable for other clones derived from the same host cell lines.
微载体提供了较大的黏附面积，可使生物反应器系统中的细胞密度较高。本研究聚焦于通过考虑细胞动力学和生理学数据，调查在四种不同微载体（Biosilon®、Microhex®、Cytodex 3®、Cytoline 2®）上培养的贴壁依赖性CHO-K1细胞系和非贴壁依赖性Ag8骨髓瘤细胞系的细胞黏附及细胞生长特性。实验在静态和搅拌细胞培养系统中进行，先使用24孔组织培养板，然后使用50毫升转瓶。在搅拌培养中，对于CHO-K1细胞系和Ag8细胞系，分别使用Cytodex 3®和Cytoline 2®微载体时获得了最高比生长速率（CHO-K1细胞系为0.026 h，Ag8细胞系为0.061 h）。代谢特性在使用四种微载体的培养物之间表现出一些差异。最显著的是，与Ag8细胞相比，CHO-K1细胞使用微载体时乳酸产量更高。扫描电子显微镜分析揭示了细胞与微载体一起时形态上的差异。在Cytodex 3®和Cytoline 2®上，CHO-K1细胞通过长而细的丝状伪足附着于基质，而在Biosilon®和Microhex®上，细胞通过覆盖基质呈现扁平形态。对于所有类型的微载体，Ag8细胞在整个培养过程中都保持球形。为了扩大规模，在50毫升转瓶中进行生产。评估了Cytodex 3®（用于CHO-K1细胞）和Cytoline 2®（用于Ag8细胞）。结果表明，通过将细胞固定在每个培养系统中可以实现高生物量产量。微载体上的细胞培养，特别是在Cytodex 3®和Cytoline 2®上的培养，分别代表了用于CHO-K1和Ag8大规模培养的良好微载体潜力。此外，由于细胞系和培养基是特定的，结果将适用于源自相同宿主细胞系的其他克隆。
