Neonatal porcine pancreatic cell clusters as a potential source for transplantation in humans: characterization of proliferation, apoptosis, xenoantigen expression and gene delivery with recombinant AAV.
Neonatal porcine islets are characterized by reproducible isolation success and high yields, sizable advantages over adult islets. In this work we have analyzed selected phenotypic and functional characteristics of porcine neonatal islets relevant to their possible use for transplant in humans. We show that porcine islet cells proliferate in culture, and synthesize and store islet-specific hormones. Proliferating beta cells can be easily identified. Implant of cultured neonatal islets in immunodeficient rodents results in the reversal of diabetes, albeit with delay. We also show that measurable apoptosis occurs in cultured neonatal porcine islets. Further, antigens recognized by human natural antibodies are expressed in a dynamic fashion over the culture period analyzed and are not limited to the alpha-Gal epitope. Lastly, we demonstrate that a recombinant Adeno-Associated virus can be used to efficiently deliver a reporter gene in porcine islets. This characterization might be helpful in the definition of the potential use of neonatal porcine islets for human transplantation.
Vizzardelli
Caterina
C
Diabetes Research Institute, Cell Transplant Center, University of Miami School of Medicine, FL 33136, USA.
Molano
R Damaris
RD
Pileggi
Antonello
A
Berney
Thierry
T
Cattan
Pierre
P
Fenjves
Elizabeth S
ES
Peel
Alyson
A
Fraker
Chris
C
Ricordi
Camillo
C
Inverardi
Luca
L
D000818:Animals; D000831:Animals, Newborn; D000910:Antibodies, Heterophile / Q000032:analysis; D015478:Antigens, Heterophile / Q000032:analysis*; D017209:Apoptosis / Q000502:physiology*; D001973:Bromodeoxyuridine; D002455:Cell Division; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology / Q000502:physiology*; D016381:Islets of Langerhans Transplantation / Q000473:pathology / Q000502:physiology*; D018613:Microscopy, Confocal; D013552:Swine; D014183:Transplantation, Heterologous / Q000473:pathology / Q000502:physiology*
D000910:Antibodies, Heterophile; D015478:Antigens, Heterophile; D007328:Insulin; D001973:Bromodeoxyuridine