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

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

Cells (2022-03-26)
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Fetal Bovine Serum, USA origin, Heat Inactivated, sterile-filtered, suitable for cell culture, suitable for insect cell culture, suitable for hybridoma
Sigma-Aldrich
SB 202190 monohydrochloride hydrate, ≥98% (HPLC)
Sigma-Aldrich
AC16 Human Cardiomyocyte Cell Line, AC16 Human Cardiomyocytes can be serially passaged and can differentiate when cultured in mitogen-free medium. The cells may be used to study developmental regulation of cardiomyocytes.