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Updated: Aug 15, 2025

Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media
Published on: May 13, 2022
Isolation of extracellular vesicles from bitch's amnion-derived cells culture and their CD59 expression: Preliminary
S Pastore1, A Troisi2, R Romani3
1Department of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126, Perugia, Italy.
Abstract:
Extracellular vesicles (EVs) are small spherical particles surrounded by a membrane with an unusual lipid composition and a striking cholesterol/phospholipidic ratio. About 2000 lipid and 3500 protein species were identified in EVs secreted by different cell sources. EVs mediate cell to cell communication in proximity to or distant from the cell of origin. In particular, it was suggested that they represent modulators of multiple processes during pregnancy. The aim of this study was to identify the presence of EVs in canine amnion-derived cells (ASCs) culture and the expression of CD 59 on their surface. Amniotic membrane was collected in PBS with antibiotics added from 2 bitches during elective caesarean section. Cells culture was prepared and EVs were isolated. EVs were used to evaluate CD59 expression by flow cytofluorimetry. We found that the majority of EVs expressed CD59. Our results could increase the knowledge about the complex mechanisms that regulate the pregnancy in the bitch.

