Isolation and characterization of placental stem cells

JULIETA L. MAYMÓ; ANTONIO PÉREZ-PÉREZ; MARTA MAGATTI; BERNARDO MASKIN; ORNELLA PAROLINI; VÍCTOR SÁNCHEZ-MARGALET; CECILIA L. VARONE

Madrid

Congreso; XXXVI Congreso de la Sociedad Española de Bioquímica y Biología Molecular; 2013

?Isolation and characterization of placental stem cells? Julieta L. Maymóa, Antonio Pérez Pérezb, Marta Magattic, Bernardo Maskind, Ornella Parolinic, Víctor Sánchez-Margaletb and Cecilia L. Varonea   aDepto. de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires- IQUIBICEN-CONICET, Buenos Aires, Argentina. bDepto. de Bioquímica Médica y Biología Molecular, Universidad de Sevilla, Sevilla, España. CCentro di Ricerca E. Menni- Fondazione Poliambulanza- Istituto Ospedaliero, Brescia, Italia. dHospital Nacional Alejandro Posadas, Buenos Aires, Argentina.   The placenta and fetal membranes have recently been proposed as an important stem cells source for regenerative medicine. Gestational  tissues offer considerable advantages over other stem cells such as bone marrow or embryo-derived cells. There is a virtually unlimited potential supply of, and easy access to such tissues, and minimal ethical and legal barriers are associated with their collection and use. Placental-derived stem cells also have the advantage of being obtained without the need for an invasive procedure. Two types of cells can be isolated from the amnios of the human placenta at term: ephitelial amniotic cells (hAECs) and mesenchymal amniotic cells (hAMCs). Both express stem cells markers and have the ability to differentiate toward all three germ layers. The hAMSCs have a fibroblast morphology, growth in colonies and posses immunomodulatory properties. The hAECs lack telomerase expression, are non tumorigenic and exhibit cobblestone appearance in culture. These properties, the simple isolation and the easy availability of the placenta, suggest that the amnios might be a useful and non controversial source of stem or progenitor cells for transplantation and regenerative medicine. The aim of this work was the isolation and characterization of human amniotic epithelial cells. We have successfully isolated amniotic cells by a specific protocol, reaching a 70% of vitality and 90% of purity, measured by flow citometry analysis. This purity was confirmed by microscopy. We have also observed cells morphology during two weeks. We determined by Western blot the activation of  MAPK and PI3K, two signaling pathways related to cellular growth, proliferation and differentiation. We observed the stimulation of ERK 1/2 and Akt phosphorylation, after 24 h serum deprivation. In summary, we have achieved an efficient isolation and extended culture of placental stem cells, with enough quantity to begin subsequent molecular, cellular and preclinical studies