Effect of Mare's age and recovery methods on the recovery rate of equine follicular oocytes for IVM procedures

Erice, I.; Gil, L.; Josa, A.; Echegaray, A.; Martínez, Francisco José Martínez; Espinosa, E.

doi:10.1016/s0093-691x(98)00022-3

Cited by 4 publications

(3 citation statements)

References 9 publications

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“…Therefore, measures taken to facilitate oocyte retrieval and to enhance oocyte maturation processes are important. In the present study, the total number of accessible follicles per ovary was similar to those reported by Erice et al (1998), higher than that of other studies (Del Campo et al 1992) but lower than those showed by Wesson and Ginther (1981) and Driancourt et al (1982). These later authors used serial histological sections in small-size follicles (less than 5 mm in diameter) which, according to a recent work (Hinrichs and Schmidt 2000), have a low meiotic competence.…”

Section: Discussionsupporting

confidence: 80%

Equine oocyte maturation with epidermal growth factor

Lorenzo

Liu

Carneiro

et al. 2002

Equine Veterinary Journal

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Summary Epidermal growth factor (EGF) has been shown to have a positive effect during oocyte in vitro maturation in several species. This study was performed to establish the capacity of equine oocytes to undergo nuclear maturation in the presence of EGF and to localise its receptor in the equine ovary by immunohistochemical methods. Oocytes were obtained by aspiration and subsequent scraping from equine follicles (15–25 mm diameter) and cultured in 3 different treatment groups for 36 h: control Group (modified TCM 199 with 0.003% BSA), EGF Group (TCM‐199 supplemented with 50 ng/ml EGF) and EMS Group (TCM 199 supplemented with 10% v/v oestrous mare serum). Each group was divided further into 3 treatments with tyrphostin A‐47, a specific tyrosine kinase inhibitor, at 0, 10−4 and 10−6 mmol/l. Maturation was determined as the percentage of oocytes reaching metaphase II stage at the end of the culture period. Immunohistochemical detection of EGF‐receptor (EGFR) was performed using a streptoavidinbiotin method. The recovery rate and oocyte retrieval were 84.6% (recovered oocytes/follicles aspirated) and 6.55 (oocytes/mare), respectively. Treatment with EGF significantly (P<0.05) increased the incidence of metaphase II stage compared with the control group (69.4 vs. 26.9% in controls, respectively). The specific‐tyrosine kinase inhibitor A‐47 was effective in suppressing EGF‐effect on EGF‐cultured oocytes; no significant differences were observed in EMS‐supplemented oocytes when cultured with A‐47. EGF‐receptor was localised in follicles, with localisation being more prominent in the cumulus than in mural granulosa cells. This finding, together with the increase of oocyte nuclear maturation rate when using EGF in culture media and the inhibition of maturation by tyrphostin A‐47, suggests a physiological role for EGF in the regulation of equine oocyte maturation. The results should help successful development of assisted reproductive technology in the horse.

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Section: Discussionsupporting

confidence: 80%

Equine oocyte maturation with epidermal growth factor

Lorenzo

Liu

Carneiro

et al. 2002

Equine Veterinary Journal

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“…Accordingly, Erice et al (1998) performed the same technique using 20G and 25G needles and 2 and 5 ml syringes, obtaining an average of 1.8 oocytes/ovary. The sample amount used in this study to evaluate the follicular wall scraping technique was considered satisfactory, considering that other studies have reported similar numbers of ovaries (311 to 517 ovaries) (CURCIO et al, 2006;CREMONESI et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Recovery of equine oocytes by scraping of the follicular wall with different specifications of needles and morphological analysis of cumulus oophorus

et al. 2015

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In follicular aspiration, physical aspects are of high significance for the technique to succeed, such as vacuum pressure, caliber of the needle and the way the follicular wall curettage is performed. The aim of this study was to investigate the recovery rate of equine oocytes aspirated by scraping of the follicular wall, testing different calibers of disposable needles, as well as the morphological evaluation of the cumulus oophorus complexes (COCs). Mares ovaries (n=447) obtained at a local slaughterhouse were transported to the laboratory in a thermal container (20 °C) and had the tunica albuginea and connective tissues dissected. The aspirated follicles had 10 to 25 mm in diameter, and 30x8 (21G 1 ¼) or 40x12 (18G 1 ½) needles were used for the aspiration, forming group A (G-A) and group B (G-B), respectively. In G-A and G-B, 480 and 548 follicles were aspirated, respectively. Under the stereomicroscope, the oocytes were evaluated according to the quality of the ooplasm and characteristics of the cumulus cells (grade I, II, III and denuded). The statistical analysis was performed using the Student's t-test, logistic regression and test of proportions, and differences were considered significant when P<0.05. There was no difference between recovery rates of groups G-A (66.5%; 330/496) and G-B (65.5%; 359/548 ResumoNa obtenção de oócitos para a espécie equina, aspectos físicos são de alta significância para o sucesso da técnica, como a pressão de vácuo e o calibre de agulha utilizado, além da forma como é realizada a raspagem da parede folicular. O objetivo deste estudo foi investigar o índice de recuperação de oócitos equinos aspirados pela técnica de raspagem da parede folicular, testando calibres distintos de agulhas descartáveis, assim como avaliação morfológica dos complexos cumulus oophorus (CCOs). Foram utilizados ovários de éguas (n=447), obtidos em abatedouro local, transportados ao laboratório em recipiente térmico (20 ºC) e submetidos à dissecação da túnica albugínea e tecidos conectores. Os folículos aspirados obtinham diâmetro entre 10 mm a 25 mm, e para tanto utilizou-se a agulha 30x8 (21G 1 ¼) ou 40x12 (18G 1 ½), formando respectivamente, o grupo A (G-A) e grupo B (G-B). No G-A e G-B aspiraram-se 480 e 548 foliculos, respectivamente. Sob lupa estéreo-microscópica avaliou-se os oócitos quanto à qualidade do ooplasma e características das células do cumulus oophorus (grau I, II, III e desnudo). Para análise estatística utilizou-se teste t Student, regressão logistica e teste de proporções, e as diferenças foram consideradas significativas quando P<0,05. A taxa de recuperação entre G-A e G-B não apresentou diferença; 66,5% (330/496)

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“…The experiments were usually based on slaughterhouse material; however, recently the transvaginal ultrasound-guided puncture of follicles has become more popular in the horse (Goudet et al 1997;Hinrichs and Williams 1997). The recovery rates for equine oocytes are very low when compared to cattle or pig, varying between 1.0 and 3.7 cells per ovary depending on the collection method used (Willis et al 1991;Shabpareh et al 1993;Alm and Torner 1994;DelCampo et al 1995;Sosnowski et al 1997;Erice et al 1998).…”

Section: Introductionmentioning

confidence: 99%