Protocol for the Ovine Amniotic Epithelial Cell In Vitro Culture and Differentiation

Hashimita Sanyal*, Annunziata Mauro, Valentina Russo, Paolo Berardinelli, Angelo Canciello, Maura Turriani and Barbara Barboni

Received: June 28, 2018;   Published: July 13, 2018

DOI: 10.26717/BJSTR.2018.06.001400

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Abstract

Ovine Amniotic epithelial cells (oAEC) are a subject of placental stem cells with great regenerative and immunomodulatory properties. Indeed, oAEC are object of intense study for regenerative medicine thanks to the several advantages in developing pre-clinical studies on a high value translational animal model, such as sheep. For this reason, a critical standardization of in vitro culture practices is fundamental in order to maintain during amplification the oAEC native phenotype, improving both oAEC in vivo therapeutic potential and clinical outcomes. Here, is described an oAEC culture protocol with supplementation of Progesterone and Estradiol hormones, able to modulate the native epithelial phenotype during the in vitro amplification. In addition, it is described the culture protocol that is able to differentiate oAECs towards osteogenic and chondrogenic lineage.

Keywords: Amniotic Epithelial Cells; Progesterone; Estradiol; Osteogenic Differentiation; Chondrogenic Differentiation; Cell Culture; Stem Cells; In Vitro Amplification

Abbreviations: OAEC: Ovine Amniotic Epithelial Cells; AM: Amniotic Membrane; PBMC: Peripheral Blood Mononuclear Cell; EMT: Epithelial-Mesenchymal Transition; FBS: Fetal Bovine Serum; BSA: Bovine Serum Albumin; α-MEM: Alpha Minimum Essential Eagle Medium; TGF-β1: Transforming Growth Factor-beta 1; PBS: Phosphate Buffered Saline; DMSO: Dimethyl Sulphoxide; EDTA: Ethylene Diaminete-tracetic Acid; RT: Room Temperature; MFI: Mean Fluorescence Intensity; DPI: Individual Protection Devices

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