In vitro and in vivo cardiomyogenic differentiation of amniotic fluid stem cells.
Cell therapy has developed as a complementary treatment for myocardial regeneration. While both autologous and allogeneic uses have been advocated, the ideal candidate has not been identified yet. Amniotic fluid-derived stem (AFS) cells are potentially a promising resource for cell therapy and tissue engineering of myocardial injuries. However, no information is available regarding their use in an allogeneic context. c-kit-sorted, GFP-positive rat AFS (GFP-rAFS) cells and neonatal rat cardiomyocytes (rCMs) were characterized by cytocentrifugation and flow cytometry for the expression of mesenchymal, embryonic and cell lineage-specific antigens. The activation of the myocardial gene program in GFP-rAFS cells was induced by co-culture with rCMs. The stem cell differentiation was evaluated using immunofluorescence, RT-PCR and single cell electrophysiology. The in vivo potential of Endorem-labeled GFP-rAFS cells for myocardial repair was studied by transplantation in the heart of animals with ischemia/reperfusion injury (I/R), monitored by magnetic resonance imaging (MRI). Three weeks after injection a small number of GFP-rAFS cells acquired an endothelial or smooth muscle phenotype and to a lesser extent CMs. Despite the low GFP-rAFS cells count in the heart, there was still an improvement of ejection fraction as measured by MRI. rAFS cells have the in vitro propensity to acquire a cardiomyogenic phenotype and to preserve cardiac function, even if their potential may be limited by poor survival in an allogeneic setting.
Stem Cell Processing Laboratory-Fondazione Città della Speranza, Venetian Institute of Molecular Medicine (VIMM), University of Padua, Via G. Orus, 2, 35129, Padua, Italy. s.bollini@ich.ucl.ac.uk
Sveva
S
Bollini
Michela
M
Pozzobon
Muriel
M
Nobles
Johannes
J
Riegler
Xuebin
X
Dong
Martina
M
Piccoli
Angela
A
Chiavegato
Anthony N
AN
Price
Marco
M
Ghionzoli
King K
KK
Cheung
Anna
A
Cabrelle
Paul R
PR
O'Mahoney
Emanuele
E
Cozzi
Saverio
S
Sartore
Andrew
A
Tinker
Mark F
MF
Lythgoe
Paolo
P
De Coppi
D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D054338:Cell Transdifferentiation; D018920:Coculture Techniques; D004195:Disease Models, Animal; D005260:Female; D005434:Flow Cytometry; D006084:Graft Rejection; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015428:Myocardial Reperfusion Injury / Q000628:therapy; D009206:Myocardium / Q000473:pathology; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology; D018995:Myosin Heavy Chains / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D059010:Single-Cell Analysis; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology; D014184:Transplantation, Homologous; D019210:Troponin I / Q000378:metabolism
D000943:Antigens, Differentiation; D011993:Recombinant Fusion Proteins; D019210:Troponin I; D049452:Green Fluorescent Proteins; D018995:Myosin Heavy Chains