Microstructured multi-well plate for three-dimensional packed cell seeding and hepatocyte cell culture.
In this article, we present a microstructured multi-well plate for enabling three-dimensional (3D) high density seeding and culture of cells through the use of a standard laboratory centrifuge to promote and maintain 3D tissue-like cellular morphology and cell-specific functionality in vitro without the addition of animal derived or synthetic matrices or coagulants. Each well has microfeatures on the bottom that are comprised of a series of ditches/open microchannels. The dimensions of the microchannels promote and maintain 3D tissue-like cellular morphology and cell-specific functionality in vitro. After cell seeding with a standard pipette, the microstructured multi-well plates were centrifuged to tightly pack cells inside the ditches in order to enhance cell-cell interactions and induce formation of 3D cellular structures during cell culture. Cell-cell interactions were optimized based on cell packing by considering dimensions of the ditches/open microchannels, orientation of the microstructured multi-well plate during centrifugation, cell seeding density, and the centrifugal force and time. With the optimized cell packing conditions, we demonstrated that after 7 days of cell culture, primary human hepatocytes adhered tightly together to form cord-like structures that resembled 3D tissue-like cellular architecture. Importantly, cell membrane polarity was restored without the addition of animal derived or synthetic matrices or coagulants.
Science and Technology, Corning Incorporated , Corning, New York 14831-0001, USA.
Vasiliy N
VN
Goral
Massachusetts General Hospital, Harvard Medical School, Charlestown , Massachusetts 02129, USA.
Sam H
SH
Au
Science and Technology, Corning Incorporated , Corning, New York 14831-0001, USA.
Ronald A
RA
Faris
Science and Technology, Corning Incorporated , Corning, New York 14831-0001, USA.
Po Ki
PK
Yuen