title: Human Cardiac Progenitor Spheroids Exhibit Enhanced Engraftment Potential.
abstract: A major obstacle to an effective myocardium stem cell therapy has always been the delivery and survival of implanted stem cells in the heart. Better engraftment can be achieved if cells are administered as cell aggregates, which maintain their extra-cellular matrix (ECM). We have generated spheroid aggregates in less than 24 h by seeding human cardiac progenitor cells (hCPCs) onto methylcellulose hydrogel-coated microwells. Cells within spheroids maintained the expression of stemness/mesenchymal and ECM markers, growth factors and their cognate receptors, cardiac commitment factors, and metalloproteases, as detected by immunofluorescence, q-RT-PCR and immunoarray, and expressed a higher, but regulated, telomerase activity. Compared to cells in monolayers, 3D spheroids secreted also bFGF and showed MMP2 activity. When spheroids were seeded on culture plates, the cells quickly migrated, displaying an increased wound healing ability with or without pharmacological modulation, and reached confluence at a higher rate than cells from conventional monolayers. When spheroids were injected in the heart wall of healthy mice, some cells migrated from the spheroids, engrafted, and remained detectable for at least 1 week after transplantation, while, when the same amount of cells was injected as suspension, no cells were detectable three days after injection. Cells from spheroids displayed the same engraftment capability when they were injected in cardiotoxin-injured myocardium. Our study shows that spherical in vivo ready-to-implant scaffold-less aggregates of hCPCs able to engraft also in the hostile environment of an injured myocardium can be produced with an economic, easy and fast protocol.
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Oltolina
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Zamperone
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Donato
authors.initials: D
authors.lastname: Colangelo
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Gregoletto
authors.affiliation: Dept. Translational Medicine, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Reano
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Pietronave
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Merlin
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Talmon
authors.affiliation: Dept. of Cardiac Surgery, Clinica S. Gaudenzio, Novara, Italy.
authors.forename: Eugenio
authors.initials: E
authors.lastname: Novelli
authors.affiliation: Dept. of Cardiac Surgery, Clinica S. Gaudenzio, Novara, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Diena
authors.affiliation: Institute Pasteur Cenci-Bolognetti, DAHFMO, Roma, Italy; Unit of Histology and Medical Embryology, IIM, Sapienza University of Rome, Rome, Italy.
authors.forename: Carmine
authors.initials: C
authors.lastname: Nicoletti
authors.affiliation: Institute Pasteur Cenci-Bolognetti, DAHFMO, Roma, Italy; Unit of Histology and Medical Embryology, IIM, Sapienza University of Rome, Rome, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Musarò
authors.affiliation: Dept. Translational Medicine, Università del Piemonte Orientale "A. Avogadro", Novara, Italy.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Filigheddu
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy; Centro di Biotecnologie per la Ricerca Medica Applicata (BRMA), Novara, Italy.
authors.forename: Antonia
authors.initials: A
authors.lastname: Follenzi
authors.affiliation: Dept. Health Sciences, Università del Piemonte Orientale "A. Avogadro", Novara, Italy; Centro di Biotecnologie per la Ricerca Medica Applicata (BRMA), Novara, Italy.
authors.forename: Maria
authors.initials: M
authors.lastname: Prat
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D005260:Female; D005455:Fluorescent Antibody Technique; D006321:Heart / Q000502:physiology*; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018874:Spheroids, Cellular / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D012333:RNA, Messenger