Extracellular matrix from decellularized mesenchymal stem cells improves cardiac gene expressions and oxidative resistance in cardiac C-kit cells

Abstract

Objective Myocardial infarction remains the number one killer disease worldwide. Cellular therapy using cardiac c-kit cells (CCs) are capable of regenerating injured heart. Previous studies showed mesenchymal stem cell-derived (MSC) extracellular matrices can provide structural support and are capable of regulating stem cell functions and differentiation. This study aimed to evaluate the effects of human MSC-derived matrices for CC growth and differentiation. Methods Human Wharton's Jelly-derived MSCs were cultured in ascorbic acid supplemented medium for 14 days prior to decellularisation using two methods. 1% SDS/Triton X-100 (ST) or 20 mM ammonia/Triton X-100 (AT). CCs isolated from 4-week-old C57/BL6N mice were cultured on the decellularised MSC matrices, and induced to differentiate into cardiomyocytes in cardiogenic medium for 21 days. Cardiac differentiation was assessed by immunocytochemistry and qPCR. All data were analysed using ANOVA. Results In vitro decellularisation using ST method caused matrix delamination from the wells. In contrast, decellularisation using AT improved the matrix retention up to 30% (p < 0.05). This effect was further enhanced when MSCs were cultured in cardiogenic medium, with a matrix retention rate up to 90%. CCs cultured on cardiogenic MSC matrix (ECM cardio ), however, did not significantly improve its proliferation after 3 days (p < 0.05), but the viability of CCs was augmented to 67.2 ± 0.7% after 24-h exposure to H 2 O 2 stress as compared to 42.9 ± 0.5% in control CCs ( p < 0.05). Furthermore, CCs cultured on cardiogenic MSC matrices showed 1.7-fold up-regulation in cardiac troponin I (cTnI) gene expression after 21 days (p < 0.05). Conclusion Highest matrix retention can be obtained by decellularization using Ammonia/Triton-100 in 2-D culture. ECM cardio could rescue CCs from exogenous hydrogen peroxide and further upregulated the cardiac gene expressions, offering an alternate in vitro priming strategy to precondition CCs which could potentially enhance its survival and function after in vivo transplantation.

Engineering Cell‐Derived Matrices: From 3D Models to Advanced Personalized Therapies

“…[ 37 ] Further decellularization allows either the reseeding of the CDMs in patients or with other cell types, or to use them acellularly for applications in regenerative medicine. [ 38,39 ] The previously described decellularization approaches can be used to produce several micrometer thick sheets of ECM mimicking native tissue properties based on the cell source. [ 40 ] Prewitz et al.…”

“…[37] Further decellularization allows either the reseeding of the CDMs in patients or with other cell types, or to use them acellularly for applications in regenerative medicine. [38,39] The previously described decellularization approaches can be used to produce several micrometer thick sheets of ECM mimicking native tissue properties based on the cell source. [40] Prewitz et al demonstrated that mesenchymal stem cell (MSC)derived ECM coatings can act as an ex vivo niche for human hematopoietic stem cell (hHSC) expansion, generating up to a threefold increase in the number of cells compared to conventional plasma-treated tissue culture plates.…”

“…[37] Furthermore, CDM sheets can be used as patches to deliver growth factors and other molecules that can act as chemoattractants for cell recruitment in tissue regeneration. [39,[41][42][43]…”

“…CDMs from MSCs upregulate cardiac gene expression in c-kit cells, promoting their survival and functions after in vivo transplantation. [39] Research in different diseases has evolved from using 2D substrates to 3D environments to identify the mechanisms underlying diseases and their progression. We believe that CDMs are excellent platforms to model the different aspects of a disease and identify possible treatment targets, given that CDMs can mimic most of the properties of native tissues and provide cells with a complex environment that is as close as possible to in vivo conditions.…”

“…CDMs from MSCs upregulate cardiac gene expression in c‐kit cells, promoting their survival and functions after in vivo transplantation. [ 39 ]…”