Determination of the appropriate concentration of sodium alginate used for in vitro culture of cat preantral follicles in a serum-free medium containing FSH, EGF and IGF-I

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dc.contributor.authorFontanillas Pérez, Juan Carlos
dc.contributor.authorGonzález Gil, Alfredo
dc.contributor.authorGarcía-Cuenca Ariati, Isabel
dc.contributor.authorPicazo González, María Dolores
dc.contributor.authorFuertes Recuero, Manuel
dc.date.accessioned2024-02-02T10:41:26Z
dc.date.available2024-02-02T10:41:26Z
dc.date.issued2023
dc.descriptionAuthor contributions M. Fuertes-Recuero: Preantral follicle isolation and culture, hormone analyses, image analyses and writing the manuscript. A. González-Gil: Design of experiments, preantral follicle isolation and culture, image analyses, writing and critical review of manuscript. J.C. Fontanillas Pérez: Hormone analyses and critical review of manuscript. I. García-Cuenca Ariati: Hormone analyses and critical review of manuscript. R.A. Picazo: Design of experiments, preantral follicle isolation and culture, image analyses, writing, critical review of manuscript and funding search.
dc.description.abstractCulture of domestic cat preantral follicles can be a suitable technology to assist oocyte conservation strategies in the family Felidae. This research was aimed to comparatively analyse cat preantral follicular development of follicles directly seeded on growth surface or encapsulated in 0.5 or 1% of sodium alginate in a serum-free medium containing FSH, EGF and IGF-I. Preantral follicles were isolated from cat ovarian cortical tissue after ovariectomy. Alginate was dissolved at 0.5 or 1% in PBS. Follicles, 4 per well, with 0% (G-0%), 0.5% (G-0.5%) or 1% (G-1%) of sodium alginate were cultured in M199 with FSH (100 ng/mL), EGF (100 ng/mL) and IGF-I (100 ng/mL) for 7 days at 37°C, 5% CO2 and 99% humidity. Culture medium was replaced every 48 h and samples were stored at −20°C until ELISA of steroid hormones. Morphometric evaluation of follicles was performed every 24 h. G-0% follicles showed granulosa cell migration away from the oocyte and disrupted morphology, whereby they reached apparently larger diameters (203.70 ± 5.82 μm; p < .05) than G-0.5% and G-1% follicles (157.89 ± 8.47 μm and 95.23 ± 1.67 μm, respectively) which maintained three-dimensional organization, being larger in G-0.5% than in G-1% (p < .05). G-0.5% follicles attained the multi-layer preantral follicle stage on day 7 of culture, whereas G-1% follicles underwent progressive atresia. On day 6, steroid concentrations were higher (p < .05) in G-0% than in G-1%: 60 ± 19 vs 0.88 ± 0.32 pg/mL oestradiol; 2.6 ± 0.84 vs 0.04 ± 0.02 ng/mL progesterone; 1.3 ± 0.22 vs 0.61 ± 0.04 ng/mL testosterone and 1.6 ± 0.54 vs 0.22 ± 0.07 ng/mL androstenedione respectively. Steroid concentrations in G-0.5% were comprised between those of G-0% and G-1% (p > .05). In conclusion, two-layer cat preantral follicles encapsulated in 0.5% alginate cultured in medium containing FSH, EGF and IGF-I can develop up to the multi-layer preantral stage in 7 days of culture, whereas follicles directly seeded on growth surface or encapsulated in 1% alginate lost their three-dimensional organization, and experienced regression with compromised steroidogenesis, respectively.
dc.description.departmentDepto. de Fisiología
dc.description.facultyFac. de Veterinaria
dc.description.refereedTRUE
dc.description.sponsorshipUniversidad Complutense de Madrid
dc.description.statuspub
dc.identifier.citationFuertes‐Recuero, M., et al. «Determination of the Appropriate Concentration of Sodium Alginate Used for in Vitro Culture of Cat Preantral Follicles in a Serum‐free Medium Containing FSH , EGF and IGF‐I». Reproduction in Domestic Animals, vol. 58, n.o 5, mayo de 2023, pp. 670-78. DOI.org (Crossref), https://doi.org/10.1111/rda.14336.
dc.identifier.doi10.1111/rda.14336
dc.identifier.essn1439-0531
dc.identifier.issn0936-6768
dc.identifier.officialurlhttps://doi.org/10.1111/rda.14336
dc.identifier.relatedurlhttps://onlinelibrary.wiley.com/doi/10.1111/rda.14336
dc.identifier.urihttps://hdl.handle.net/20.500.14352/98168
dc.issue.number5
dc.journal.titleReproduction in Domestic Animals
dc.language.isoeng
dc.page.final678
dc.page.initial670
dc.publisherWiley
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internationalen
dc.rights.accessRightsopen access
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.cdu636.09
dc.subject.keywordAlginate
dc.subject.keywordCat
dc.subject.keywordPreantral follicle
dc.subject.keywordSteroid hormones
dc.subject.ucmVeterinaria
dc.subject.unesco3109 Ciencias Veterinarias
dc.titleDetermination of the appropriate concentration of sodium alginate used for in vitro culture of cat preantral follicles in a serum-free medium containing FSH, EGF and IGF-I
dc.title.alternativeDeterminación de la concentración apropiada de sodioalginato utilizado para el cultivo in vitro de folículos preantrales de gato en unsuero-medio libre que contiene FSH, EGF e IGF-I
dc.typejournal article
dc.type.hasVersionVoR
dc.volume.number58
dspace.entity.typePublication
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