Punción folicular 289 PUNCIÓN FOLICULAR (OVUM PICK-UP, OPU) EN LA VACA

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1 Punción folicular 289 PUNCIÓN FOLICULAR (OVUM PICK-UP, OPU) EN LA VACA Peter Bols Introducción Aunque un objetivo de interés es conocer los factores que afectan la recolección post-mortem del ovocito, la punción de los folículos ováricos inmaduros es particularmente interesante desde el punto de vista biotecnológico, cuando se lleva a cabo en animales donantes vivos. En ese sentido, los ovocitos obtenidos son destinados a la producción in vitro de embriones durante un prolongado período. Lo cual obviamente no es posible con donantes faenadas o sacrificadas. La recolección de ovocitos por medio de la punción folicular de hembras vivas también permite la modulación endocrina de la actividad ovárica de la donante para influir sobre la cantidad y calidad de los complejos ovocito cumulus (COC) recuperados. Mientras la recuperación de los ovocitos, después de la muerte, se hace simplemente aspirando los folículos ováricos con una jeringa y una aguja, es diferente cuando se aspiran los folículos inmaduros directamente del ovario en una donante viva. La literatura es bastante escasa sobre los efectos del procedimiento de aspiración sobre el número, la morfología y la capacidad de desarrollo subsecuente del complejo recolectado. Existen diferencias hipotéticas básicas entre la aspiración post-mortem e in vivo. En primer lugar, la manipulación transrectal del ovario es necesaria para la recuperación del ovocito en la hembra donante viva, por medio de la visualización del folículo en forma indirecta, a través de laparoscopia, conocida también como imágen de ultrasonido (imaging ultrasound), ver más abajo. En contraste con ello, cuando se puncionan los folículos de ovarios de matadero, el operador puede examinar el ovario directa y completamente. Así, un folículo específico puede escogerse y puede puncionarse bajo control visual directo. Segundo, se puede esperar un efecto mecánico diferente que puede desempeñar un rol importante, mientras el folíuclo es puncionado in vitro con una jeringa y aguja, comparado con la aspiración folicular in vivo, con una presión de vacío de aspiración ajustable, como fue demostrado por Hashimoto y col. (1999a). Finalmente, el uso de hembras donantes vivas permite obtener información adicional sobre el momento de recolección del ovocito, como también conocer los valores endocrinos de la sangre, para ganar más información sobre el estado metabólico de la donante y su posible efecto sobre la calidad del ovocito. Los factores principales que controlan el resultado, con respecto al número y calidad de los ovocitos recuperados al puncionar los folículos inmaduros por medio de OPU en la vaca, pueden ser divididos en 2 categorías principales (Bols, 1997, 2005). La primera categoría incluye los aspectos más técnicos: el procedimiento de aspiración, tipo de aguja y diámetros, más la posible influencia de la presión de vacío de aspiración sobre la morfología de los complejos (COCs). Para documentar este aspecto técnico, se llevaron a cabo varios estudios, examinando el desarrollo y probando la aguja del dispositivo de OPU, que se comentará más adelante. Una segunda categoría puede describirse como aquellos factores biológicos, que incluyen, entre muchos otros, el estímulo hormonal previo a la punción folícular, el momento en que se lleva a cabo el procedimiento en relación con el estado puntual del ciclo estral, la edad, la raza y la condición fisiológica de la donante. De hecho, la recolección de los ovocytos de donantes vivas debe ser considerada como un proceso dinámico. Cuando los ovocitos son recolectados luego de un estímulo hormonal externo, debe esperarse una influencia consecuente sobre la recolección y la calidad de los ovocitos. El capítulo presentará una descripción actualizada de la metodología de la aspiración folicular, el equipamiento necesario y los resultados de la recolección de los ovocitos en hembras donantes vivas.

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9 Punción folicular 297 Scott CA, Robertson L, de Moura RTD, Paterson C, JS Boyd (1994) Technical aspects of transvaginal ultrasound-guided follicular aspiration in cows Vet Rec, Seneda MM, Esper CS, Garcia JM, de Oliveira JA, R Vantini (2001) Relationship between follicle size and ultrasound-guided transvaginal oocyte recovery Anim Reprod Sci, Sheldon IM, H Dobson (2003) Reproductive challenges facing the cattle industry at the beginning of the 21 st century Reproduction Supplement, 1-13 Shioya Y, Kuwayama M, Fukushima M, Iwasaki S, A Hanada (1988) In vitro fertilization and cleavage capability of bovine follicular oocytes classified by cumulus cells and matured in vitro Theriogenology, Simon L, Bungartz L, Rath D, H Niemann (1993) Repeated bovine oocyte collection by means of a permanently rinsed ultrasound guided aspiration unit Theriogenology, 312 Singh J, Adams GP, RA Pierson (2003) Promise of new imaging technologies for assessing ovarian function Anim Rep Sci, Sirois J, JE Fortune (1988) Ovarian follicular dynamics during the estrous cycle in heifers monitored by real time ultrasonography Biol Reprod, Sosnowski J, Switonski M, Lechniak D, K Molinski (1996) Cytogenetic evaluation of in vitro matured bovine oocytes collected from ovaries of individual donors Theriogenology, Staigmiller RB, RM Moor (1984)Effect of follicle cells on the maturation and developmental competence of ovine oocytes matured outside the follicle Gamete Research, Stubbings RB, JS Walton (1995) Effect of ultrasonically-guided follicle aspiration on estrous cycle and follicular dynamics in Holstein cows Theriogenology, Takagi Y, Mori K, Takahashi T, Sugawara S, Masaki J Differences in development of bovine oocytes recovered by aspiration or by mincing J Anim Sci 1992(), Taneja M, Bols PEJ, Van de Velde A, Ju J-C, Schreiber D, Tripp MW, Levine H, Echelard Y, Riesen J, X Yang (2000) Developmental competence of juvenile calf oocytes in vitro and in vivo: influence of donor animal variation and repeated gonadotropin stimulation Biol Reprod, Thonon F, Ectors FJ, Delval A, Fontes RS, Touati K, JF Beckers (1993) In vitro maturation, fertilization and development rates of bovine oocytes connected with the reproductive status of the donor Theriogenology, 330 Van Arendonck JAM, A-E Liinamo (2003) Dairy cattle production in Europe Theriogenology, van der Schans A, van der Westerlaken CAJ, de Wit AAC, Eyestone WH, HA de Boer (1991) Ultrasound-guided transvaginal collection of oocytes in the cow Theriogenology, 288 Van Soom A, Van Vlaenderen, Mahmoudzadeh AR, Ysebaert MT, A de Kruif (1994a) Salvage of oocytes from sterile genetically valuable cows, resulting in the birth of a calf Anim Reprod Sci, Van Soom A, Mijten P, Van Vlaenderen I, Van den Branden J, Mahmoudzadeh AR, A de Kruif (1994b) Birth of double-muscled Belgian Blue calves after transfer of in vitro produced embryos into dairy cattle Theriogenology, Van Soom A, Bols PEJ, A de Kruif (1995) Improved results in IVF-treatment of sterility patients by careful selection of bulls and by stimulation of cows with FSH before slaughter Reprod Dom Anim, 67 Van Wagtendonck-de Leeuw AM, Mullaart E, de Roos APW, Merton JS, den Daas JHG, Kemp B, L de Ruigh (2000) Effects of different reproduction techniques: AI, MOET or IVP, on health and welfare of bovine offspring Theriogenology, Vergos E In vitro fertilization and embryo culture in cattle PhD Thesis, 1990, National University of Ireland, Dublin Vos PLAM, de Loos FAM, Pieterse MC, Bevers MM, Taverne MAM, SJ Dieleman (1994) Evaluation of transvaginal ultrasound-guided follicle puncture to collect oocytes and follicular fluids at

10 298 Bols consecutive times relative to the preovulatory LH surge in ecg/pg treated cows Theriogenology, Ward FA, Lonergan P, Enright BP, MP Boland (2000) Factors affecting recoverey and quality of oocytes for bovine embryo production in vitro using ovum pick-up technology Theriogenology, Wikland M, Enk L, Hammarberg K, L Nilsson (1987) Use of vaginal transducer for oocyte retrieval in an IVF/ET program J Clin Ultrasound, Wurth YA, ThAM Kruip (1992) Bovine embryo production in vitro after selection of the follicles and oocytes Proceedings of the 12th International Congress on Animal Reproduction (The Hague), Zhang L, Jiang S, Wozniak PJ, Yang X, RA Godke (1995) Cumulus cell function during bovine oocyte maturation, fertilization and embryo development in vitro Mol Reprod Dev,

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