Effects of cell storage and passage on basal and oxytocin-regulated prostaglandin secretion by equine endometrial epithelial and stromal cells |
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Authors: | Szóstek A Z Siemieniuch M J Galvão A M Lukasik K Zieba D Ferreira-Dias G M Skarzynski D J |
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Institution: | a Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Olsztyn, Poland b C.I.I.S.A., Faculty of Veterinary Medicine, Technical University of Lisbon, Lisbon, Portugal c Laboratory of Biotechnology and Genomics, Department of Swine and Small Ruminant Breeding, Faculty of Animal Sciences, Agricultural University of Krakow, Poland |
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Abstract: | Cell cultures are useful for determining the responses of specific cell types to various factors under controlled conditions and for obtaining a better understanding of in vivo physiologic processes. The aims of the present study were (i) to establish methodologies for isolation, culture and cryopreservation of equine endometrial epithelial and stromal cells; and (ii) to determine the effect of passage and cryopreservation on endometrial cell physiology, based on their basal and oxytocin (OT)-stimulated prostaglandin (PG) release. Epithelial and stromal cells were obtained by enzymatic digestion of equine endometrium collected from Days 2-5 of the estrous cycle (n = 16). Primary epithelial and stromal cells, as well as cryopreserved cells were stimulated with OT (10−7m) for 24 h. The concentrations of PGE2 and PGF2α in the culture medium were measured by enzyme-linked immunosorbent assay (EIA). Oxytocin increased PGE2 and PGF2α release by primary cultures of unfrozen epithelial cells until passage I (P < 0.01) and by the primary culture of unfrozen and cryopreserved/thawed stromal cells until passage IV (P < 0.01). Cryopreserved/thawed stromal cells cultured up to passage IV and unfrozen epithelial cells derived from passage I have physiological properties similar to those observed in primary culture and may be successfully used for in vitro studies of PG secretion. |
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Keywords: | Endometrium Cryopreservation Oxytocin Prostaglandin Mare |
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