Macromolecular crowding facilitates rapid fabrication of intact, robust cell sheets

Abstract:Objectives To develop a rapid and simple method to fabricate intact, robust cell sheets from common cell culture dishes by combination of a macromolecular crowding (MMC) reagent and vitamin C. Results It was found that 3T3 fibroblasts or human bone marrow mesenchymal stem cells (hBMSCs) and their secreted cell derived extracellular matrices could be easily detached as intact cell sheets under gently pipetting after treated by MMC and vitamin C for 4 days. This method also allowed fabrication of functional multi-layered hepatic cell sheets by culturing 10×104 cells/cm2 HepG2 cells on top of confluent 3T3 fibroblast layers. What’s more, MMC induced hBMSC cell sheets demonstrated 1.4 times larger area and 1.6 times greater cell number than that of cell sheets harvested from temperature-responsive cell culture dishes. Conclusion MMC based method make it possible to fabricate various types of cell sheets more conveniently, economically, and thus may facilitate wide application of cell sheet technology.