Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes
The choice of medium and supplements can affect meiotic regulation and may have an impact on the regulation of mammalian oocyte growth and embryonic cell function. The aim of the present study was to assess the effects of oxygen concentration and endogenous lanosterol on the in vitro maturation (IVM...
| Autores principales: | , , |
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| Formato: | article |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/5564 |
| _version_ | 1855032298621108224 |
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| author | Marco-Jiménez, Francisco Vicente, José Salvador Viudes-De-Castro, María P. |
| author_browse | Marco-Jiménez, Francisco Vicente, José Salvador Viudes-De-Castro, María P. |
| author_facet | Marco-Jiménez, Francisco Vicente, José Salvador Viudes-De-Castro, María P. |
| author_sort | Marco-Jiménez, Francisco |
| collection | ReDivia |
| description | The choice of medium and supplements can affect meiotic regulation and may have an impact on the regulation of mammalian oocyte growth and embryonic cell function. The aim of the present study was to assess the effects of oxygen concentration and endogenous lanosterol on the in vitro maturation (IVM) media without serum and based on recombinant human chorionic gonadotrophin in prepubertal ewe oocytes. Firstly, the effect of varying oxygen concentrations (5% and 20%) during IVM in TCM-199 supplemented (4 mg/ml bovine serum albumin (BSA), 100 M cysteamine, 0.3 mM sodium pyruvate, 0.1 UI/ml recombinant follicle-stimulating hormone (r-FSH; Gonal-F (R) 75 UI, Serono, Italy), 0.1 UI/ml recombinant leuteinizing hormone (r-LH; Lhadi (R) 75 UI, Serono, Italy) and 1 g/ml estradiol-17) on subsequent nuclear maturation of oocytes examined under ultraviolet light following staining with bisbenzimide (Hoechst 33342) was investigated. Secondly, two concentrations of lanosterol (0, 10 and 50 M) were added to the IVM medium. Nuclear maturation of oocytes was examined as previously. Lipid content in oocytes, an important indicator of cytoplasmic maturity, was also measured using Nile red fluorescent stain. The results showed that low oxygen concentration affected the nuclear maturation. Similarly, a significantly higher rate of meiosis resumption was observed with 10 M (72.3%) of lanosterol compared with the control (51.8%) or 50 M of lanosterol (59.4%). A significantly higher content of lipids was also observed with 10 and 50 M of lanosterol (7.3 +/- 0.2 x 10(6) and 7.4 +/- 0.2 x 10(6) arbitrary units of fluorescence) compared with the control (6.7 +/- 0.2 x 10(6) arbitrary units of fluorescence). The results indicate that 10 M lanosterol during IVM in medium without serum and based on recombinant human chorionic gonadotrophin has a positive effect on maturation of prepubertal ewe oocytes. |
| format | article |
| id | ReDivia5564 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia55642025-04-25T14:43:18Z Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes Marco-Jiménez, Francisco Vicente, José Salvador Viudes-De-Castro, María P. The choice of medium and supplements can affect meiotic regulation and may have an impact on the regulation of mammalian oocyte growth and embryonic cell function. The aim of the present study was to assess the effects of oxygen concentration and endogenous lanosterol on the in vitro maturation (IVM) media without serum and based on recombinant human chorionic gonadotrophin in prepubertal ewe oocytes. Firstly, the effect of varying oxygen concentrations (5% and 20%) during IVM in TCM-199 supplemented (4 mg/ml bovine serum albumin (BSA), 100 M cysteamine, 0.3 mM sodium pyruvate, 0.1 UI/ml recombinant follicle-stimulating hormone (r-FSH; Gonal-F (R) 75 UI, Serono, Italy), 0.1 UI/ml recombinant leuteinizing hormone (r-LH; Lhadi (R) 75 UI, Serono, Italy) and 1 g/ml estradiol-17) on subsequent nuclear maturation of oocytes examined under ultraviolet light following staining with bisbenzimide (Hoechst 33342) was investigated. Secondly, two concentrations of lanosterol (0, 10 and 50 M) were added to the IVM medium. Nuclear maturation of oocytes was examined as previously. Lipid content in oocytes, an important indicator of cytoplasmic maturity, was also measured using Nile red fluorescent stain. The results showed that low oxygen concentration affected the nuclear maturation. Similarly, a significantly higher rate of meiosis resumption was observed with 10 M (72.3%) of lanosterol compared with the control (51.8%) or 50 M of lanosterol (59.4%). A significantly higher content of lipids was also observed with 10 and 50 M of lanosterol (7.3 +/- 0.2 x 10(6) and 7.4 +/- 0.2 x 10(6) arbitrary units of fluorescence) compared with the control (6.7 +/- 0.2 x 10(6) arbitrary units of fluorescence). The results indicate that 10 M lanosterol during IVM in medium without serum and based on recombinant human chorionic gonadotrophin has a positive effect on maturation of prepubertal ewe oocytes. 2017-06-01T10:12:34Z 2017-06-01T10:12:34Z 2014 FEB 2014 article Marco-Jiménez, F., Vicente, J. S., & Viudes-De-Castro, M. P. (2014). Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes. Zygote, 22(1), 50-57. 0967-1994 http://hdl.handle.net/20.500.11939/5564 10.1017/S096719941100044X en openAccess Impreso |
| spellingShingle | Marco-Jiménez, Francisco Vicente, José Salvador Viudes-De-Castro, María P. Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes |
| title | Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes |
| title_full | Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes |
| title_fullStr | Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes |
| title_full_unstemmed | Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes |
| title_short | Effect of lanosterol on the in vitro maturation in semi-defined culture system of prepubertal ewe oocytes |
| title_sort | effect of lanosterol on the in vitro maturation in semi defined culture system of prepubertal ewe oocytes |
| url | http://hdl.handle.net/20.500.11939/5564 |
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