Table 1.
Summary of cardiac tissue-derived dECM scaffolds used for producing cardiac patch for myocardial infarction treatment
| Species | Decellularization agents | Scaffold thickness | Cell types | In vitro | In vivo | Biological results | Refs |
|---|---|---|---|---|---|---|---|
| Rat | 1% SDS; 1% Triton X-100/0.5% EDTA | N/A | Human induced pluripotent stem cell-derived CMs; human induced pluripotent stem cell-derived CD90+ cells | × | × | Cardiac dECM scaffold enhanced the maturation of human iPSC-derived cardiac cells in vitro. After seeding, the dECM scaffold exhibited normal electrical properties and responded to the pharmaceutical agents. When patched on the acute rat MI model, the recellularized cardiac dECM has shown to reduce infract size, increase in wall thickness and promote vascularization | [26] |
| Rat | 10 mM Tris HCl/0.1% EDTA; 0.2% SDS/10 mM Tris HCl | N/A | Immortalized adult Lin-Sca-1+ cardiac progenitor cells (iCPCSca-1); neonatal rat CMs | × | Fetal and adult cardiac dECM scaffolds, when seeded with cardiac progenitor cells and neonatal CMs, have shown to support the viability, proliferation and migration. Compared to adult cardiac dECM, fetal scaffold has resulted better repopulation efficiency, migration and colonization rates of seeded Lin-Sca-1+ cardiac progenitor cells and neonatal rat CMs | [39] | |
| Rat | 0.25% Triton X-100/10 mmol/l NH4OH | 10 µm* | Neonatal rat CMs | × | Neonatal rat CMs, when cultured on thin cardiac dECM slices, have exhibited higher proliferation rate and increased cardiac gene and protein expressions compared to the control group | [40] | |
| Rat | 1% SDS; 1% Triton X-100 | N/A | Induced pluripotent stem cells | × | Rat cardiac dECM supported the attachment, survival, growth and differentiation of iPSCs as indicated by decreased pluripotency markers after 7 days of culture | [41] | |
| Rat | 1% SDS; 1% Triton X-100 | 381±157 µm | N/A | × | The rat cardiac dECM had significantly higher stiffness compared to the native myocardium tissue | [42] | |
| Mouse | 0.25% SDS/0.5 mg/ml DNase | N/A | Mouse embryonic ventricular cells; mouse ESC-derived progenitors | × | The recellularization of embryonic cardiac dECM with mouse embryonic ventricular cells and mouse ESC-derived progenitors (day 5 after inducing differentiation) has resulted ESC differentiation as characterized by endothelial, cardiac and smooth muscle markers, leading to achieve beating cardiac patch after 20 h and 24 days of culture, respectively | [27] | |
| Mouse | 0.05% Trypsin/0.02% EDTA, 1.1% NaCl and 0.7% NaCl; 0.1% SDS; 1% Triton X-100 | 300 µm | Murine ESCs | × | Cardiac dECM scaffold, when seeded with murine ESCs using hanging drop method, has shown to form cell aggregates which attached, survived, proliferated and merged with adjacent aggregates during 16 days of culture | [31] | |
| Porcine and rat | SDS; Triton X-100 | 300 µm* | Neonatal rat ventricular cells | × | Neonatal rat ventricular cells, when seeded on rat or pig engineered heart slices, have promoted cell elongation, alignment and synchronous contraction, leading to the production of an anisotropic and functional tissue that could be electrically paced for electrophysiological studies | [43] | |
| Porcine | 1% Triton X-100, 1% SDS and 0.5% Trypsin | 2000 µm* | Rat myocardial fibroblast; rat neonatal CMs | × | Compared to decellularization with Trypsin and Triton X-100, the SDS-based treatment has resulted better decellularization efficiency of porcine myocardium tissue as demonstrated by complete removal of cells and better preservation of ECM microstructures. When seeded on obtained dECM scaffolds, rMFs showed distinct cell attachment and growth rate response while rCMs were different in terms of morphologies and spontaneous beating magnitudes based on the decellularization methods. | [44] | |
| Porcine | 1.1% NaCl/0.02% EDTA and 0.7% NaCl/0.02% EDTA; 0.05% Trypsin/0.02% EDTA; 1% Triton X-100 and 0.1% ammonium hydroxide | 1500 µm* | N/A | × | Decellularized porcine myocardium patch, when implanted on acute and chronic rat MI models, has shown to promote robust vascularization after implantation, recruit cardiac progenitor (GATA4+, c-kit+) and myocyte (MYLC+, TRPI+) on the patch and induce constructive ECM remodeling as indicated by increased M2/M1 macrophage phenotypic ratio, leading to the significant improvement of cardiac function | [28] | |
| Porcine | 0.1% SDS/0.01% Trypsin, 1 mM phenylmethylsulfonylfluoride and 20 µg/ml RNase A/0.2 mg/ml DNase | 2000 µm* | Porcine bone marrow mononuclear cells | × | Porcine cardiac dECM, when cultured with the mixture of undifferentiated and differentiated bone marrow mononuclear cells toward cardiac phenotype, has supported cell attachment, viability, infiltration and proliferation of seeded cells, leading to maintain the CM-like phenotype and possible endothelialization within the scaffold. | [45] | |
| Porcine | 0.1% SDS/0.01% Trypsin, 1 mM phenylmethylsulfonylfluoride and 20 µg/ml RNase A/0.2 mg/ml DNase | 2270 ± 380 µm | N/A | × | Porcine myocardium tissue, when treated with 0.1% SDS and 0.01% trypsin solution using a frame-pin supporting system and a rotating bioreactor, has resulted removal of cells, DNA fragments (∼98%) and α-Gal porcine antigens. Compared to native porcine myocardium tissue, dECM scaffold showed stiffer tensile properties | [46] | |
| Porcine | 0.1% SDS/0.01% Trypsin, 1 mM phenylmethylsulfonylfluoride and 20 µg/ml RNase A/0.2 mg/ml DNase | 3000 µm* | Rat MSCs | × | Rat MSCs seeded on porcine dECM scaffold with a needle injection, when cultured using the CM differentiation growth medium containing 5-azacytidine and subjected to mechanical and electrical stimulations (20% strain; 5 V, 1 Hz), resulted in the differentiation of MSCs toward CM-like phenotype | [47] | |
| Porcine | 10 mM Tris/0.1% EDTA; 0.5% SDS; DMEM containing 10% fetal bovine serum; 0.1% peracetic acid/4% ethanol | 150 µm* | Neonatal rat ventricular myocytes; human ESC-derived CMs; human induced pluripotent stem cell-derived CMs | × | When seeded with NRVMs, the laser-cut thin sheet of decellularized cardiac slices resulted synchronously beating and exhibited a striated pattern of organized sarcomeres. Decellularized cardiac ECM slices seeded with hESC-CMs produced beating scaffolds with measurable intracellular calcium transients and maximum twitch stress of 1.7 N/mm2. Similarly, hiPSC-CM seeded dECM slices achieved maximum peak stress of 6.5 mN/mm2 and twitch kinetics similar to the reported values from adult human trabeculae | [48] | |
| Porcine | 1.1% NaCl and 0.7% NaCl; 0.05% Trypsin/0.02% EDTA; 1% Triton X-100/1% ammonium hydroxide | 14 600 ± 19 00 µm | Rat MSCs; human umbilical vein endothelial cells | × | When compared the decellularization protocols, Triton X-100/trypsin-based perfusion method was found to be more effective to achieve thicker dECM while retaining structural characteristics, inherent vasculature, fiber morphology and mechanical properties. The thick dECM scaffold supported the attachment and long-term cell survival of rMSCs. HUVECs were found to form a monolayer surrounding the inner lumen of the inherent vasculature on dECM | [30] | |
| Porcine | 1.1% NaCl and 0.7% NaCl; 0.05% Trypsin/0.02% EDTA; 1% Triton X-100/1% ammonium hydroxide | 15 000 µm* | Bone marrow-derived MSCs; human umbilical vein endothelial cells; human ESC-derived CMs | × | Thick porcine myocardium dECM, when co-cultured with hMSCs and HUVECs under dynamic culture conditions using a perfusion bioreactor chamber, has supported compartmentalized recellularization and higher cell infiltration compared to static culture conditions, leading to functional vascularization/angiogenesis as indicated by sprouting of capillary-like vessels within the areas of scaffold containing high hMSCs (up to 1.7 mm thickness). Human ESC-derived CMs were seeded on thick dECM scaffold, and the scaffold supported CM phenotype, leading to synchronous beating 3 days after initial cell seeding | [49] | |
| Porcine | 1.1% NaCl/0.02% EDTA and 0.7% NaCl/0.02% EDTA; 0.05% Trypsin/0.02% EDTA; 1% Triton X-100 and 0.1% ammonium hydroxide | 3000 µm* | Sheep cardiac fibroblast; rat cardiac myocytes; rat bone marrow-derived MSCs | × | Cardiac fibroblast seeded on porcine dECM resulted the scaffold shrinkage and ECM remodeling, as demonstrated by significant increase of GAG content (∼23%) and ECM remodeling-related mRNAs including collagen I and III, matrix metalloproteinase 2 and type 1 tissue inhibitor of metalloproteinase as compared to control. Decellularized ECM scaffold seeded with CMs began to beat few days after initial seeding and showed positive expression for functional cardiac markers. dECM seeded with MSCs maintained cell viability over 24 days in culture | [50] | |
| Porcine | 0.02% Trypsin/0.05% EDTA/0.05% NaN3; 3% Triton X-100/0.05% EDTA/0.05% NaN3; 4% deoxycholic acid | N/A | Chicken embryonic CMs | × | Perfusion-based decellularization of porcine WH resulted cardiac dECM with well-preserved collagen, elastin, and GAGs, and mechanical integrity. Cardiac dECM sheet supported the formation of organized chicken CM sarcomere structure in vitro, as indicated by α-actinin staining for striations fibers | [25] | |
| Porcine | 0.02% Trypsin/0.05% EDTA/0.05% NaN3; 3% Triton X-100/0.05% EDTA/0.05% NaN3; 4% deoxycholic acid | 2500 µm | N/A | × | Porcine cardiac dECM patch, when used to treat right ventricular outflow tract (RVOT) defect in a Lewis rat model, has resulted CM recruitment, dECM patch remodeling and neovascularization, leading to the improved heart function | [51] | |
| Porcine | 1% SDS; 0.01% Triton X-100 | 300, 600 and 900 µm | hMSCs; rASCs | × | When cultured on top of the scaffold from one side (lateral cell seeding), decellularized porcine myocardial slices (dPMSs) supported the cell attachment with high viability and induced endothelial differentiation and maturation of hMSCs and rASCs. Compared to lateral seeding, bilateral cell seeding (cells were seeded from both sides of the scaffold) has significantly enhanced seeding efficiency, infiltration and growth in 600 μm dPMS | [52] | |
| Porcine | 1% SDS; 0.01% Triton X-100 | 300 µm | rASCs; pig adipose-derived stem cells | × | × | Rat and pig ASCs, when seeded on 300 μm dPMS, has shown the distinct responses in terms of cell attachment, viability, infiltration and proliferation. The rASCs cultured on dPMS showed endothelial differentiation. When used to deliver rASCs using dPMS on a rat MI model, a higher number of transplanted cells were present in the infracted area compared to direct injection seeding method, leading to increased vascular formation within the patch | [38] |
| Human and porcine | 10 mM Tris/0.1% EDTA; 0.5% SDS | 300 µm* | Human umbilical cord blood-derived MSCs; murine iPSC-derived CMs; murine neonatal CMs | × | Cardiac dECM scaffold promoted cell attachment, viability and proliferation of human umbilical cord blood-derived MSCs, murine iPSC-derived CMs and murine neonatal CMs. Compared to MSCs, iPSC-derived CMs showed less cell attachment, proliferation and infiltration on the human dECM slices | [29] | |
| Human | 1% SDS | N/A | Human cardiac progenitor cells; hMSCs; human umbilical-vein endothelial cells; H9c2 rat CMs; HL-1 CMs | × | When seeded on human dECM scaffold, hCPCs has shown to enhance the expression of cardiac markers including bMHC, MEF2C, Nkx2.5 and TnnT. HUVECs cultured on dECM formed a lining of endocardium and vasculature. The CMs organized into nascent muscles bundles and showed mature calcium dynamics and electrical coupling on dECM scaffold | [24] | |
| Human | 1% SDS; 1% Triton X-100 | 200 µm | Human iPSC-derived CMs | × | Human iPSC-derived CMs, when seeded on human dECM slices, has supported the cell attachment, viability and function as evidenced by spontaneously contracting slices within 4–7 days of culture and the presence of sarcomeric structure, cell-mediated matrix deformation, electrical conduction and contractile force | [36] | |
| Human | 10 mM Tris/0.1% EDTA; 0.5% SDS | 300 µm* | Murine ESCs; murine induced pluripotent stem cells; murine mesenchymal stromal cells | × | Human cardiac tissue-derived dECM favored attachment, viability, proliferation, and cardiac lineage commitment of seeded ESCs and iPSCs as evidenced by positive immunohistochemistry staining for cardiac troponin T and heavy-chain cardiac myosin as well as significant increase of mRNA expression for myosin heavy polypeptide 6, cardiac troponin T2 and NK2 homeobox 5. MSCs showed no evidence of CM differentiation | [32] | |
| Human |
|
350 µm* | Human cardiac primitive cells | × | Human cardiac primitive cells, when cultured on dECM scaffold, has promoted the differentiation of seeded cells toward CMs and smooth muscle cells as indicated by distinct gene expression for CMs (MEF2C, ACTC1) and smooth muscle cells (GATA6, ACTA2) | [53] | |
| Human | 1% SDS | 400 µm | Human ESC-derived CM-like cells; human induced pluripotent stem cell-derived CM-like cells | × | Human cardiac dECM scaffold, when cultured with hPSC-derived CLCs, has shown to promote the differentiation and maturation toward CMs as demonstrated by enhanced electrophysiological properties and positive immunofluorescence staining for alpha-sarcomeric actinin, Troponin T, MYH6, NKX2.5 and CX43 after 10 days of culture | [54] |
The asterisk sign (*) denotes the thickness of dECM before tissue decellularization. For these studies, the scaffold thickness after decellularization procedure has not been reported. N/A indicates not applicable.