3D In Vitro Platform for Cell and Explant Culture in Liquid-like Solids

用于在类液态固体中培养细胞和外植体的3D体外平台

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作者:Duy T Nguyen ,Jack E Famiglietti ,Ryan A Smolchek ,Zadia Dupee ,Nickolas Diodati ,Diego I Pedro ,Juan M Urueña ,Matthew A Schaller ,W Gregory Sawyer

Abstract

Existing 3D cell models and technologies have offered tools to elevate cell culture to a more physiologically relevant dimension. One mechanism to maintain cells cultured in 3D is by means of perfusion. However, existing perfusion technologies for cell culture require complex electronic components, intricate tubing networks, or specific laboratory protocols for each application. We have developed a cell culture platform that simply employs a pump-free suction device to enable controlled perfusion of cell culture media through a bed of granular microgels and removal of cell-secreted metabolic waste. We demonstrated the versatile application of the platform by culturing single cells and keeping tissue microexplants viable for an extended period. The human cardiomyocyte AC16 cell line cultured in our platform revealed rapid cellular spheroid formation after 48 h and ~90% viability by day 7. Notably, we were able to culture gut microexplants for more than 2 weeks as demonstrated by immunofluorescent viability assay and prolonged contractility.

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