To this day, the concept of continuous bioprocessing has been applied mostly to the manufacture of molecular biologics such as proteins, growth factors, and secondary metabolites with biopharmaceutical uses. The present work now sets to explore the potential application of continuous bioprocess methods to source large numbers of human adherent cells with potential therapeutic value. To this purpose, we developed a smart multifunctional surface coating capable of controlling the attachment, proliferation, and subsequent self-detachment of human corneal stromal cells. This system allowed the maintenance of cell cultures under steady-state growth conditions, where self-detaching cells were continuously replenished by the proliferation of those remaining attached. This facilitated a closed, continuous bioprocessing platform with recovery of approximately 1% of the total adherent cells per hour, a yield rate that was maintained for 1 month. Moreover, both attached and self-detached cells were shown to retain their original phenotype. Together, these results represent the proof-of-concept for a new high-throughput, high-standard, and low-cost biomanufacturing strategy with multiple potentials and important downstream applications.

中文翻译：

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开发用于基质细胞制造的连续生物加工方法

时至今日，连续生物加工的概念主要应用于分子生物制品的制造，例如蛋白质、生长因子和具有生物制药用途的次级代谢物。目前的工作现在着手探索连续生物过程方法的潜在应用，以获取大量具有潜在治疗价值的人体贴壁细胞。为此，我们开发了一种智能多功能表面涂层，能够控制人角膜基质细胞的附着、增殖和随后的自脱离。该系统允许在稳态生长条件下维持细胞培养，其中自分离细​​胞通过剩余附着细胞的增殖不断补充。这促进了一个封闭的、连续生物处理平台，每小时可回收约 1% 的总贴壁细胞，产量可维持 1 个月。此外，附着细胞和自分离细胞均显示保留其原始表型。总之，这些结果代表了具有多种潜力和重要下游应用的新的高通量、高标准和低成本生物制造策略的概念验证。