建立和优化 AMBR15 高通量模拟灌流模型。
Establishment and optimization of a high-throughput mimic perfusion model in ambr 15.
机构信息
School of Life Science and Technology, China Pharmaceutical University, #639 Longmian Dadao, Jiangning District, Nanjing, 211198, Jiangsu, People's Republic of China.
Cell Culture Process Development Department, WuXi Biologics, #288 Fute Middle Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, People's Republic of China.
出版信息
Biotechnol Lett. 2021 Feb;43(2):423-433. doi: 10.1007/s10529-020-03026-5. Epub 2020 Nov 13.
OBJECTIVES
To establish an automated high-throughput mimic perfusion scale-down model (SDM) in ambr 15 system.
RESULTS
An optimized SDM for mimic perfusion was developed in ambr 15 system. Cell retention in ambr 15 was realized by sedimentation and supernatant removal with a retention rate > 95%. Although the SDM couldn't reach the viable cell density (VCD) at a bench scale bioreactor (BR), it maintained VCD at approximately 30 × 10 cells/mL with a cell bleeding rate estimated theoretically and predicted the cell specific perfusion rate (CSPR). A base-feeding strategy was developed to alleviate the pH drop during sedimentation which would adversely have an impact on cell growth, and showed an apparent cell viability improvement from 79.6% (control) to 90.1% on Day 18. The optimized SDM for mimic perfusion was employed for media screening in two cell lines.
CONCLUSIONS
A small-scale high-throughput perfusion model in ambr 15 was developed, optimized to improve cell viability, and as a result, utilized for media screening in two cell lines.
目的
在 ambr15 系统中建立自动化高通量模拟灌流缩小模型（SDM）。
结果
在 ambr15 系统中开发了用于模拟灌流的优化 SDM。通过沉降和上清液去除实现了 ambr15 中的细胞保留，保留率>95%。尽管 SDM 无法达到生物反应器（BR）的工作规模的活细胞密度（VCD），但它通过理论估计和预测细胞比灌流率（CSPR）来维持约 30×10 个细胞/mL 的 VCD。开发了基础进料策略来缓解沉降过程中的 pH 值下降，这会对细胞生长产生不利影响，并在第 18 天显示出细胞活力从 79.6%（对照）明显提高到 90.1%。优化的模拟灌流 SDM 用于两种细胞系的培养基筛选。
结论
在 ambr15 中开发了小型高通量灌注模型，对其进行了优化以提高细胞活力，并最终用于两种细胞系的培养基筛选。
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