Mesenchymal Stem Cell Seeding of Porcine Small Intestinal Submucosal Extracellular Matrix for Cardiovascular Applications

Abstract

In this study, we investigate the translational potential of a novel combined construct using an FDA-approved decellularized porcine small intestinal submucosa extracellular matrix (SIS-ECM) seeded with human or porcine mesenchymal stem cells (MSCs) for cardiovascular indications. With the emerging success of individual component in various clinical applications, the combination of SIS-ECM with MSCs could provide additional therapeutic potential compared to individual components alone for cardiovascular repair. We tested the in vitro effects of MSC-seeding on SIS-ECM on resultant construct structure/function properties and MSC phenotypes. Additionally, we evaluated the ability of porcine MSCs to modulate recipient graft-specific response towards SIS-ECM in a porcine cardiac patch in vivo model. Specifically, we determined: 1) in vitro loading-capacity of human MSCs on SIS-ECM, 2) effect of cell seeding on SIS-ECM structure, compositions and mechanical properties, 3) effect of SIS-ECM seeding on human MSC phenotypes and differentiation potential, and 4) optimal orientation and dose of porcine MSCs seeded SIS-ECM for an in vivo cardiac application. In this study, histological structure, biochemical compositions and mechanical properties of the FDA-approved SIS-ECM biomaterial were retained following MSCs repopulation in vitro. Similarly, the cellular phenotypes and differentiation potential of MSCs were preserved following seeding on SIS-ECM. In a porcine in vivo patch study, the presence of porcine MSCs on SIS-ECM significantly reduced adaptive T cell response regardless of cell dose and orientation compared to SIS-ECM alone. These findings substantiate the clinical translational potential of combined SIS-ECM seeded with MSCs as a promising therapeutic candidate for cardiac applications.

Conditioned medium: a new alternative for cryopreservation of equine umbilical cord mesenchymal stem cells

“…Although it has disadvantages mainly related to immunogenicity for clinical application purposes, FBS has an important role in the cryopreservation process by stabilizing cell membranes, protecting cells from free radicals and adjusting osmotic pressure (Marquez‐Curtis et al, ). Many studies, used FBS for cultivation (Barberini et al, ; Maia et al, ; Chang et al, ; Tang et al, ) and cryopreservation (Mambell et al, ; Martinello et al, ; Hennes et al, ; Kim et al, ; Munévar et al, ; Duan and Lopez, ; Ercolin et al, ; Irioda et al, ) possibly by its positive aspect in cell growth and benefits during freezing. However, it is important to point that for therapeutic purposes, the FBS must be washed out before cell transfer (Maia et al, ).…”

“…Although it has disadvantages mainly related to immunogenicity for clinical application purposes, FBS has an important role in the cryopreservation process by stabilizing cell membranes, protecting cells from free radicals and adjusting osmotic pressure (Marquez-Curtis et al, 2015). Many studies, used FBS for cultivation (Barberini et al, 2014;Maia et al, 2015;Chang et al, 2016;Tang et al, 2016) and cryopreservation (Mambell et al, 2009;Martinello et al, 2010;Hennes et al, 2015;Figure 3 Median and interquartile range (IQR) of cell viability (a), necrosis (b), early apoptosis (c), and late apoptosis þ necrosis (d) of equine umbilical cord intervascular matrix mesenchymal stem cells (UCIM-MSCs) by flow cytometry before (fresh) and after cryopreservation with medium M1-M4. Asterisks ( Ã ) denote significant differences (P < 0.05) between the fresh cells and treatments (cryopreservation medium: M1-M4).…”

Biomaterial and stem cell‐based strategies for skeletal muscle regeneration

“…Immunogenic concerns have been reported with decellularized scaffolds, poly(ethylene glycol), and hyaluronic acid based materials. Immunomodulation via immunosuppressant drugs or paracrine action of stem cells may be able to regulate the intensity and duration of macrophage phenotypes.…”

Tissue and organ decellularization in regenerative medicine

“…Seeding autologous oral mucosa epithelial cells along with SIS resulted in muscular regeneration and re‐epithelization after 8 weeks of surgery and the results were superior to SIS with no cells . Applying Food and Drug Administration (FDA)‐approved SIS ECM seeded with mesenchymal stem cells resulted in a lower response of the adaptive immune system in a porcine model, when compared with SIS–ECM with no cells …”

“…47 Applying Food and Drug Administration (FDA)-approved SIS ECM seeded with mesenchymal stem cells resulted in a lower response of the adaptive immune system in a porcine model, when compared with SIS-ECM with no cells. 48…”