Cell Sheet Technology for Cardiac Tissue Engineering

Abstract

In this chapter, we describe the methods for the fabrication and transfer/transplantation of 3D tissues by using cell sheet technology for cardiac tissue regeneration. A temperature-responsive culture surface can be fabricated by grafting a temperature-responsive polymer, poly(N-isopropylacrylamide), onto a polystyrene cell culture surface. Cells cultured confluently on such a culture surface can be recovered as an intact cell sheet, and functional three-dimensional (3D) tissues can then be easily fabricated by layering the recovered cell sheets without any scaffolds or complicated manipulation. Cardiac cell sheets, myoblast sheets, mesenchymal stem cell sheets, cardiac progenitor cell sheets, etc., which are prepared from temperature-responsive culture surfaces, can be easily transplanted onto heart tissues of animal models, and those cell sheet constructs enhance the cell transplant efficiency, resulting in the induction of effective therapy.

Three-Dimensional Human Cardiac Tissue Engineered by Centrifugation of Stacked Cell Sheets and Cross-Sectional Observation of Its Synchronous Beatings by Optical Coherence Tomography

“…After detachment, the cardiac cell sheet with medium was transferred onto a polystyrene culture dish and was spread out on the dish by three manipulations using a pipette: (i) rotation of the dish, (ii) slow aspiration, and/or (iii) gentle dropping of culture medium as shown in previous reports [5, 23], and thereafter, the medium was removed to promote attachment between the cell sheet and the culture surface (Figure 1(a)). Just after the transfer, many spaces were detected between them by OCT, even after the medium had been removed (Figure 2(a-1)).…”

“…Connexin 43 was also detected at the edge of neighboring cardiomyocytes in a human iPS cell-derived cardiac cell sheet, and the electrical coupling of the cardiac cell sheet was shown by an electrophysiological analysis using the extracellular potential detection method [17, 33]. Additionally, the synchronous beatings and electrical coupling of layered cardiac cell sheets, which were fabricated by human iPS cell-derived cardiac cells or mouse embryonic stem (ES) cell-derived cardiac cells, were shown by the using of (i) phase-contrast microscopy, (ii) the electrophysiological analysis, and (iii) an intracellular calcium transient imaging method [17, 23, 34]. In this study, the rapid electrical coupling of layered human iPS cell-derived cardiac cell sheets, which were fabricated rapidly by centrifugation, was suggested by the cross-sectional observation of OCT.…”

Rapid creation system of morphologically and functionally communicative three‐dimensional cell‐dense tissue by centrifugation

“…The tissues created within a PDMS ring device were able to be easily transferred to another surface by a transfer device (Supplementary Movie 2), showing the created tissues were transplantable even immediately after the harvest. A 3D tissue can be transferred onto target tissues including intensively beating heart tissue of a porcine model by using the transfer device . Due to the innate/biological characteristics of cells, like the reaggregation ability and tissue‐reconstruction ability, conventional scaffold‐free tissue engineering methodology requires several days to create a 3D tissue.…”

“…Human induced pluripotent stem (iPS) cells [201B7 stain (RBRC‐HPS0001)] were provided by the RIKEN BRC through the National Bio‐Resources Project of the MEXT, Japan. Human iPS cells and C2C12 mouse myoblasts (Sumitomo Dainippon Pharma, Osaka, Japan) were cultured, and human iPS cell‐derived cardiac cells were obtained as described in our previous studies . Two types of temperature‐responsive culture surfaces (24‐well culture plate and 60‐mm culture dish) (UpCell® plate/dish, CellSeed Inc., Tokyo, Japan) were used for creating detachable 3D tissues.…”

From cardiac tissue engineering to heart-on-a-chip: beating challenges

“…The first concept focuses on the generation of sheets of confluent cells which may then be either directly used or further stacked together into multi-layer structures, the so called cell-sheet engineering [44, 45] (Fig. 2A).…”