Tissue Eng Regen Med.  2021 Oct;18(5):807-818. 10.1007/s13770-021-00363-y.

Multilayered Cell Sheets of Cardiac Reprogrammed Cells for the Evaluation of Drug Cytotoxicity

• Kwon SP
• Song SY
• Yoo J
• Kim HY
• Lee JR
• Kang M
• Sohn HS
• Go S
• Jung M
• Hong J
• Lim S
• Kim C
• Moon S
• Char K
• Kim BS

• KMID: 2520639
• DOI: http://doi.org/10.1007/s13770-021-00363-y

Abstract

BACKGROUND
Various cell-culture systems have been used to evaluate drug toxicity in vitro. However, factors that affect cytotoxicity outcomes in drug toxicity evaluation systems remain elusive. In this study, we used multilayered sheets of cardiac-mimetic cells, which were reprogrammed from human fibroblasts, to investigate the effects of the layer number on drug cytotoxicity outcomes.
METHODS
Cell sheets of cardiac-mimetic cells were fabricated by reprogramming of human fibroblasts into cardiacmimetic cells via coculture with cardiac cells and electric stimulation, as previously described. Double-layered cell sheets were prepared by stacking the cell sheets. The mono- and double-layered cell sheets were treated with 5-fluorouracil (5-FU), an anticancer drug, in vitro. Subsequently, apoptosis and lipid peroxidation were analyzed. Furthermore, effects of cardiacmimetic cell density on cytotoxicity outcomes were evaluated by culturing cells in monolayer at various cell densities.
RESULTS
The double-layered cell sheets exhibited lower cytotoxicity in terms of apoptosis and lipid peroxidation than the mono-layered sheets at the same 5-FU dose. In addition, the double-layered cell sheets showed better preservation of mitochondrial function and plasma membrane integrity than the monolayer sheets. The lower cytotoxicity outcomes in the double-layered cell sheets may be due to the higher intercellular interactions, as the cytotoxicity of 5-FU decreased with cell density in monolayer cultures of cardiac-mimetic cells.
CONCLUSION
The layer number of cardiac-mimetic cell sheets affects drug cytotoxicity outcomes in drug toxicity tests. The in vitro. cellular configuration that more closely mimics the in vivo configuration in the evaluation systems seems to exhibit lower cytotoxicity in response to drug.

Keyword

Cardiac-mimetic cell; Cell sheet; Drug cytotoxicity; PLGA nanoporous membrane; Direct reprogramming