Peculiarity of Porcine Amniotic Membrane and Its Derived Cells: A Contribution to the Study of Cell Therapy from a Large Animal Model

Anna Lange-Consiglio, Bruna Corradetti, Sabrina Bertani, Valentina Notarstefano, Claudia Perrini, Maria Giovanna Marini, Silvana Arrighi, Giampaolo Bosi, Angelo Belloli, Davide Pravettoni, Valentina Locatelli, Fausto Cremonesi, Davide Bizzaro

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)472-483
Number of pages12
JournalCellular Reprogramming
Volume17
Issue number6
DOIs
StatePublished - Dec 1 2015

ASJC Scopus subject areas

  • Biotechnology
  • Developmental Biology
  • Cell Biology

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