title: Optimal concentration of hyaluronan and plant protein in different culture systems for in vitro maturation of bovine oocytes.
abstract: With a view to search for optimal concentration of hyaluronan (HA) and plant protein (PP) in different culture systems for in vitro maturation of bovine oocytes, cumulus-oocyte complexes (COCs) were matured in vitro in 2 culture systems (first co-cultured with granulose cells and estrus calf serum (ECS) in 2 mL volume, second without co-culture where ECS was replaced by exogenous hormones and BSA or PP in 100 microL dose under mineral oil). Seven types of media were used; 3 in first system and 4 in second system. To evaluate HA and PP effect on oocytes after in vitro culture an estimation of meiosis stage and a level of DNA fragmentation was performed by TUNEL staining. The highest meiotic maturation (84%) was observed in oocytes cultured in medium enriched with ECS in co-culture with granulose cells (1st system). The lowest meiotic maturation was noted in medium with addition of BSA (43%). The addition of HA in the medium enriched with BSA significantly increased the rate of matured oocytes (67%) and also didn't affect the chromatin quality of individual oocytes. The addition of HA to the culture medium supplemented with a PP decreased the rate of matured oocytes to 54% but no statistical differences were noted. The results of the present study showed that HA supplementation didn't have a detrimental impact on oocyte chromatin integrity and improved bovine oocytes' meiotic maturation in medium supplemented only with BSA without co-culture of granulose cells.
authors.affiliation: Department of Biotechnology of Animal Reproduction, National Research Institute of Animal Production, Krakowska 1 st., Balice/Kraków, Poland. jolanta.opiela@izoo.krakow.pe
authors.forename: Jolanta
authors.initials: J
authors.lastname: Opiela
authors.forename: Ewa
authors.initials: E
authors.lastname: Latasiewicz
authors.forename: Zdzisław
authors.initials: Z
authors.lastname: Smorag
mesh_terms: D000818:Animals; D002417:Cattle; D002469:Cell Separation; D002478:Cells, Cultured / Q000166:cytology / Q000187:drug effects; D002843:Chromatin / Q000648:ultrastructure; D018920:Coculture Techniques; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism; D006820:Hyaluronic Acid / Q000032:analysis / Q000494:pharmacology*; D020287:In Situ Nick-End Labeling; D059471:In Vitro Oocyte Maturation Techniques; D008540:Meiosis / Q000187:drug effects; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D009866:Oogenesis / Q000187:drug effects*; D010940:Plant Proteins / Q000032:analysis / Q000494:pharmacology*
chemical_list: D002843:Chromatin; D003470:Culture Media; D010940:Plant Proteins; D006820:Hyaluronic Acid