TY - JOUR
T1 - Peculiarity of Porcine Amniotic Membrane and Its Derived Cells
T2 - A Contribution to the Study of Cell Therapy from a Large Animal Model
AU - Lange-Consiglio, Anna
AU - Corradetti, Bruna
AU - Bertani, Sabrina
AU - Notarstefano, Valentina
AU - Perrini, Claudia
AU - Marini, Maria Giovanna
AU - Arrighi, Silvana
AU - Bosi, Giampaolo
AU - Belloli, Angelo
AU - Pravettoni, Davide
AU - Locatelli, Valentina
AU - Cremonesi, Fausto
AU - Bizzaro, Davide
N1 - Publisher Copyright:
© Copyright 2015, Mary Ann Liebert, Inc.
PY - 2015/12
Y1 - 2015/12
N2 - The aim of this work was to provide, for the first time, a protocol for isolation and characterization of stem cells from porcine amniotic membrane in view of their potential uses in regenerative medicine. From three samples of allanto-amnion recovered at delivery, the amniotic membrane was stripped from overlying allantois and digested with trypsin and collagenase to isolate epithelial (amniotic epithelial cells [AECs]) and mesenchymal cells, respectively. Proliferation, differentiation, and characterization studies by molecular biology and flow cytometry were performed. Histological examination revealed very few mesenchymal cells in the stromal layer, and a cellular yield of AECs of 10 × 106/gram of digested tissue was achieved. AECs readily attached to plastic culture dishes displaying typical cuboidal morphology and, although their proliferative capacity decreased to the fifth passage, AECs showed a mean doubling time of 24.77 ± 6 h and a mean frequency of one fibroblast colony-forming unit (CFU-F) for every 116.75 plated cells. AECs expressed mesenchymal stem cell (MSC) mRNA markers (CD29, CD166, CD90, CD73, CD117) and pluripotent markers (Nanog and Oct 4), whereas they were negative for CD34 and MHCII. Mesodermic, ectodermic, and endodermic differentiation was confirmed by staining and expression of specific markers. We conclude that porcine amniotic membrane can provide an attractive source of stem cells that may be a useful tool for biomedical research.
AB - The aim of this work was to provide, for the first time, a protocol for isolation and characterization of stem cells from porcine amniotic membrane in view of their potential uses in regenerative medicine. From three samples of allanto-amnion recovered at delivery, the amniotic membrane was stripped from overlying allantois and digested with trypsin and collagenase to isolate epithelial (amniotic epithelial cells [AECs]) and mesenchymal cells, respectively. Proliferation, differentiation, and characterization studies by molecular biology and flow cytometry were performed. Histological examination revealed very few mesenchymal cells in the stromal layer, and a cellular yield of AECs of 10 × 106/gram of digested tissue was achieved. AECs readily attached to plastic culture dishes displaying typical cuboidal morphology and, although their proliferative capacity decreased to the fifth passage, AECs showed a mean doubling time of 24.77 ± 6 h and a mean frequency of one fibroblast colony-forming unit (CFU-F) for every 116.75 plated cells. AECs expressed mesenchymal stem cell (MSC) mRNA markers (CD29, CD166, CD90, CD73, CD117) and pluripotent markers (Nanog and Oct 4), whereas they were negative for CD34 and MHCII. Mesodermic, ectodermic, and endodermic differentiation was confirmed by staining and expression of specific markers. We conclude that porcine amniotic membrane can provide an attractive source of stem cells that may be a useful tool for biomedical research.
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U2 - 10.1089/cell.2015.0029
DO - 10.1089/cell.2015.0029
M3 - Article
C2 - 26540004
AN - SCOPUS:84949656671
SN - 2152-4971
VL - 17
SP - 472
EP - 483
JO - Cellular Reprogramming
JF - Cellular Reprogramming
IS - 6
ER -