Development of endomyocardial fibrosis model using a cell patterning technique: In vitro interaction of cell coculture of 3T3 fibroblasts and RL-14 cardiomyocytes

Abstract

Cardiac functions can be altered by changes in the microstructure of the heart, i.e., remodeling of the cardiac tissue, which may activate pathologies such as hypertrophy, dilation, or cardiac fibrosis. Cardiac fibrosis can develop due to an excessive deposition of extracellular matrix proteins, which are products of the activation of fibroblasts. In this context, the anatomical-histological change may interfere with the functioning of the cardiac tissue, which requires specialized cells for its operation. The purpose of the present study was to determine the cellular interactions and morphological changes in cocultures of 3T3 fibroblasts and RL-14 cardiomyocytes via the generation of a platform an in vitro model. For this purpose, a platform emulating the biological characteristics of endomyocardial fibrosis was generated using a cell patterning technique to study morphological cellular changes in compact and irregular patterns of fibrosis. It was found that cellular patterns emulating the geometrical distributions of endomyocardial fibrosis generated morphological changes after interaction of the RL-14 cardiomyocytes with the 3T3 fibroblasts. Through this study, it was possible to evaluate biological characteristics such as cell proliferation, adhesion, and spatial distribution, which are directly related to the type of emulated endomyocardial fibrosis. This research concluded that fibroblasts inhibited the proliferation of cardiomyocytes via their interaction with specific microarchitectures. This behavior is consistent with the histopathological distribution of cardiac fibrosis; therefore, the platform developed in this research could be useful for the in vitro assessment of cellular microdomains. This would allow for the experimental determination of interactions with drugs, substrates, or biomaterials within the engineering of cardiac tissues.

Cardiomyocyte-targeting exosomes from sulforaphane-treated fibroblasts affords cardioprotection in infarcted rats

“…We chose the NIH/3T3 cell line for our study because it is similar to adult cardiac fibroblasts [ 33 , 60 ] and has a high mitotic rate in vitro. In addition, noncardiac fibroblasts are widely used in the production of clinical-grade exosomes for the treatment of many human diseases [ 34 ].…”

“…Mouse NIH/3T3 embryo fibroblasts (ATCC® CRL-1658™ kindly provided by Dr. Giuseppe Rainaldi, IFC-CNR, Pisa, Italy) are frequently used as in vitro models of adult cardiac fibroblasts [ 33 , 34 ] and fibroblast-cardiomyocytes exosome-based crosstalk [ 35 ]. HL-1 mouse atrial cardiomyocytes (a kind gift of Professor W. Claycomb, LSUHSC, New Orleans, LA, USA), a cell line used to study cardiac physiology at the cellular level [ 36 ] and to assess the cardioprotective effects of exosomes [ 18 , 19 , 21 , 37 , 38 ], were cultured in Claycomb Medium (Sigma-Aldrich, Merk Life Science S.r.l., Milan, Italy) [ 39 ] supplemented with 10% exosomes-depleted FBS, 100units/mL of penicillin, 100 µg/mL streptomycin, 250 ng/mL amphotericin B and 2 mM L-glutamine (Sigma-Aldrich, Merk Life Science S.r.l., Milan, Italy).…”