Cardiac fibroblast-derived VCAM-1 enhances cardiomyocyte proliferation for fabrication of bioengineered cardiac tissue

Abstract

Background Fibroblasts are indispensable for the fabrication of cell-sheet–based bioengineered cardiac tissues; however, whether cardiac fibroblasts can improve tissue properties for transplantation or in vitro models compared with other fibroblast types remains unclear. We compared the cell organization and functional properties of cardiomyocyte sheets derived from co-culture with different fibroblast types and investigated the molecular mechanisms for the observed differences. Methods and results Cardiac cell sheets were fabricated by co-culturing mouse embryonic stem cell (ESC)-derived cardiomyocytes with mouse neonatal cardiac fibroblasts (NCFs), mouse adult cardiac fibroblasts (ACFs), and mouse adult dermal fibroblasts (ADFs). Cardiac cell sheets obtained from NCF or ACF co-culture showed numerous uniformly distributed and functional (beating) cardiomyocytes, while cell sheets obtained by co-culture with ADFs showed fewer and aggregated cardiomyocytes. The greater number of cardiomyocytes in the presence of NCFs was because of enhanced cardiomyocyte proliferation, as revealed by protein markers of mitosis and BrdU incorporation. Microarray analysis revealed that NCFs expressed substantially higher levels of vascular cell adhesion molecule-1 (VCAM-1) than ADFs. Treatment of ESC-derived cardiomyocytes in monoculture with soluble VCAM-1 significantly increased the number of functional cardiomyocytes, while the enhancement of cardiomyocyte number by co-culture with NCFs was abolished by anti-VCAM-1 antibodies. Conclusions Cardiac fibroblasts enhance the proliferation of ESC-derived cardiomyocytes through VCAM-1 signaling, leading to an increase in functional myocardial cells in bioengineered tissue sheets. These sheets may be advantageous for cell-based therapy and in vitro heart research.

Human cardiac fibroblasts expressing VCAM1 improve heart function in postinfarct heart failure rat models by stimulating lymphangiogenesis

“…VCAM1 is the ligand for α4β1 integrin, which is known to exist as both a transmembrane protein and in a soluble form. Several groups, including ours, have reported that VCAM1 is expressed in cardiomyocytes [66], CSCs [2], and CFs [67]. Soluble VCAM1 secreted from murine CSCs demonstrated cardioprotective effects in mouse through the α4β1 integrin-mediated activation of Akt, ERK, and P38MAPK in cardiomyocytes [2]; this phenomenon has also been reported to be critical for heart development [68,69].…”

Construction of a vascularized hydrogel for cardiac tissue formation in a porcine model

“…It is unclear if the dermal fibroblast origin of our derived hiPSC‐CM could have played a role in such reduced remodelling capacity of fibroblasts co‐derived during cardiac differentiation. Nevertheless, dermal fibroblasts were recently reported to be less supportive of bioengineered cardiac tissue formation when compared to cardiac fibroblasts (Iwamiya, Matsuura, Masuda, Shimizu, & Okano, ).…”

Optimising platelet secretomes to deliver robust tissue‐specific regeneration

“…Our data on the composition of platelet releasate revealed that known key growth factors driving myoblast and/or chondrocyte proliferation such as PDGFBB and VEGFA were increased several fold in SR (Kieswetter, Schwartz, Alderete, Dean, & Boyan, ; Scully, Sfyri, et al, ). VCAM‐1 was previously shown to increase cardiomyocyte proliferation and skeletal myoblast differentiation and was 3.8‐fold more abundant in SR compared with physiological releasate (Choo, Canner, Vest, Thompson, & Pavlath, ; Iwamiya, Matsuura, Masuda, Shimizu, & Okano, ). Similarly, growth factors driving fibroblast proliferation such as EGF and FGF‐2 were increased 1.92‐ and 3.43‐fold, respectively (Yu, Matsuda, Takeda, Uchinuma, & Kuroyanagi, ).…”