981d2499fac9fc9e43d470de3627a726 pmc_34685551.pdf 98cfbf42488c71b9e7bee19ef6b88031b131e774 pmc_34685551.pdf 5e2dc6c0eeee766ca7bfd5a51a2d78a0fc070cbd97665084ace09dacf7698741 pmc_34685551.pdf Title: Intracoronary Delivery of Porcine Cardiac Progenitor Cells Overexpressing IGF-1 and HGF in a Pig Model of Sub-Acute Myocardial Infarction Subject: Human cardiac progenitor cells (hCPC) are considered a good candidate in cell therapy for ischemic heart disease, demonstrating capacity to improve functional recovery after myocardial infarction (MI), both in small and large preclinical animal models. However, improvements are required in terms of cell engraftment and efficacy. Based on previously published reports, insulin-growth factor 1 (IGF-1) and hepatocyte growth factor (HGF) have demonstrated substantial cardioprotective, repair and regeneration activities, so they are good candidates to be evaluated in large animal model of MI. We have validated porcine cardiac progenitor cells (pCPC) and lentiviral vectors to overexpress IGF-1 (co-expressing eGFP) and HGF (co-expressing mCherry). pCPC were transduced and IGF1-eGFPpos and HGF-mCherrypos populations were purified by cell sorting and further expanded. Overexpression of IGF-1 has a limited impact on pCPC expression profile, whereas results indicated that pCPC-HGF-mCherry cultures could be counter selecting high expresser cells. In addition, pCPC-IGF1-eGFP showed a higher cardiogenic response, evaluated in co-cultures with decellularized extracellular matrix, compared with native pCPC or pCPC-HGF-mCherry. In vivo intracoronary co-administration of pCPC-IGF1-eGFP and pCPC-HFG-mCherry (1:1; 40 106/animal), one week after the induction of an MI model in swine, revealed no significant improvement in cardiac function. Keywords: cardiac progenitor cell; IGF-1; HGF; lentiviral vectors; decellularized extracellular matrix; porcine large animal model; myocardial infarction Author: Cristina Prat-Vidal, Verónica Crisóstomo, Isabel Moscoso, Claudia Báez-Díaz, Virginia Blanco-Blázquez, Guadalupe Gómez-Mauricio, Guillermo Albericio, Susana Aguilar, María-Eugenia Fernández-Santos, Francisco Fernández-Avilés, Francisco M. Sánchez-Margallo, Antoni Bayes-Genis and Antonio Bernad Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Sep 28 11:37:45 2021 CEST ModDate: Tue Sep 28 11:47:04 2021 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 25 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4873906 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HFUITD+VnURWPalladioL Type 1 Custom yes yes no 10 0 JEERPS+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 IQBHLF+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 GJLLLM+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 YFFKLH+CMSY10 Type 1 Builtin yes yes no 28 0 JYPLJJ+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 PFKOHU+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 40 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom 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PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 613 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Cell Culture Destination: subsection.2.1 Item: Title: Karyotyping and Cell Proliferation Studies Destination: subsection.2.2 Item: Title: Flow Cytometry Destination: subsection.2.3 Item: Title: Cardiac In Vitro Differentiation of pCPC Destination: subsection.2.4 Item: Title: Quantitative Real-Time PCR Destination: subsection.2.5 Item: Title: RNA-Seq Analysis Destination: subsection.2.6 Item: Title: pCPC Ientiviral Transduction In Vitro Destination: subsection.2.7 Item: Title: Western Blot Destination: subsection.2.8 Item: Title: Immunofluorescence Destination: subsection.2.9 Item: Title: Co-Culture of Engineered pCPC in Decellularized Rat Cardiac Scaffolds Destination: subsection.2.10 Item: Title: Transplantation of Engineered pCPC in Swine Myocardial Infarct Model Destination: subsection.2.11 Item: Title: Statistical Analysis Destination: subsection.2.12 Item: Title: Results Destination: section.3 Children: Item: Title: Characterization of Adult pCPC Destination: subsection.3.1 Item: Title: Generation and Characterization of pCPC with Forced IGF-1 or HGF Overexpression Destination: subsection.3.2 Item: Title: Evaluation of the Potential Effect of IGF-1 and HGF Overexpression on pCPC Gene Expression Profile Destination: subsection.3.3 Item: Title: IGF-1 Synergizes with Decellularized Rat Cardiac Scaffolds in the Promotion of Cardiogenic Differentiation of pCPC Destination: subsection.3.4 Item: Title: In Vivo Evaluation of pCPC-IGF1-eGFP and pCPC-HGF-mCherry Co-Administration for the Treatment of Swine Myocardial Infarct Destination: subsection.3.5 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: Intracoronary Delivery of Porcine Cardiac Progenitor Cells Overexpressing IGF-1 and HGF in a Pig Model of Sub-Acute Myocardial Infarction Author: Cristina Prat-Vidal, Verónica Crisóstomo, Isabel Moscoso, Claudia Báez-Díaz, Virginia Blanco-Blázquez, Guadalupe Gómez-Mauricio, Guillermo Albericio, Susana Aguilar, María-Eugenia Fernández-Santos, Francisco Fernández-Avilés, Francisco M. Sánchez-Margallo, Antoni Bayes-Genis and Antonio Bernad Subject: Human cardiac progenitor cells (hCPC) are considered a good candidate in cell therapy for ischemic heart disease, demonstrating capacity to improve functional recovery after myocardial infarction (MI), both in small and large preclinical animal models. However, improvements are required in terms of cell engraftment and efficacy. Based on previously published reports, insulin-growth factor 1 (IGF-1) and hepatocyte growth factor (HGF) have demonstrated substantial cardioprotective, repair and regeneration activities, so they are good candidates to be evaluated in large animal model of MI. We have validated porcine cardiac progenitor cells (pCPC) and lentiviral vectors to overexpress IGF-1 (co-expressing eGFP) and HGF (co-expressing mCherry). pCPC were transduced and IGF1-eGFPpos and HGF-mCherrypos populations were purified by cell sorting and further expanded. Overexpression of IGF-1 has a limited impact on pCPC expression profile, whereas results indicated that pCPC-HGF-mCherry cultures could be counter selecting high expresser cells. In addition, pCPC-IGF1-eGFP showed a higher cardiogenic response, evaluated in co-cultures with decellularized extracellular matrix, compared with native pCPC or pCPC-HGF-mCherry. In vivo intracoronary co-administration of pCPC-IGF1-eGFP and pCPC-HFG-mCherry (1:1; 40 106/animal), one week after the induction of an MI model in swine, revealed no significant improvement in cardiac function. 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