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. 2017 Jul 27;7:6703. doi: 10.1038/s41598-017-06949-8

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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The perfusion culture setup using the spiral microfluidic cell retention device. (a) The cell culture was continuously processed through the device by a peristaltic pump. The majority of the cells were recycled back to the bioreactor while the harvest was collected in a separate container. The cell culture medium was supplied to the bioreactor using another peristaltic pump. The working volume of the bioreactor was controlled automatically based on bioreactor weight using customized software. The pH and dissolved oxygen levels were monitored and controlled to meet established set points. (b) The microfluidic cell retention device and the fluidic adaptor. (c) The bottom view of the cell retention device. The four spiral channels were combined in parallel. Scale bar, 10 mm. (d) The pre-sterilized cell retention device, tubings, and bottles were connected to each other in a sterilized manner in the biosafety cabinet. (e) The setup was transferred into the standard CO₂ incubator, and the perfusion culture began after four days.