title: In vitro embryo production in llamas (Lama glama) from in vivo matured oocytes with raw semen processed with Androcoll-E using defined embryo culture media.
abstract: The aim of this study was to carry out in vitro fertilization using spermatozoa selected with Androcoll-E™ and to evaluate the efficiency of the culture medium DMEM-F12 for in vitro embryo development in the llama. Twelve adult females from 18 superstimulated (67%) were used as oocyte donors. They were superstimulated with 1500 IU of eCG and after 5 days, received a single dose of buserelin. Twenty hours post-injection, follicular aspiration was conducted by flank laparotomy. Semen collections were performed under general anesthesia by electroejaculation of the male. The ejaculates were processed with a solution of collagenase (0.1%) and an Androcoll-E™ column was used to improve the sample. Sixty nine COCs were recovered from 79 aspirated follicles (87% recovery). Only expanded COCs were used (n = 67); they were randomly placed in groups of 1-5 in Fertil-TALP and the sperm suspension (20 × 10(6) live spermatozoa/ml) was added to each fertilization microdroplet. After 24 h, they were randomly placed in one of two culture media: SOF (n = 34) or DMEM-F12 (n = 33) and incubated for 6 days in humidified atmosphere of 5% CO(2) , 5% O(2) and 90% N(2) at 38°C. The blastocyst rate was 20% (7/34) in SOF medium (3 hatched, 2 expanded and 2 early blastocysts) and 15% (5/33) in DMEM medium (all expanded blastocysts). In conclusion, using Androcoll-E™ it is possible to select good quality spermatozoa from llama ejaculates for in vitro fertilization and to produce blastocysts in DMEM-F12 medium. This is also the first time that hatched llama blastocysts have been produced after culture in a defined medium such as SOFaa.
authors: Cátedra de Teriogenología, Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Universidad de Buenos Aires (UBA), Argentina.
authors: V
authors: V
authors: Trasorras
authors: S
authors: S
authors: Giuliano
authors: G
authors: G
authors: Chaves
authors: D
authors: D
authors: Neild
authors: A
authors: A
authors: Agüero
authors: M
authors: M
authors: Carretero
authors: M
authors: M
authors: Pinto
authors: C
authors: C
authors: Baca Castex
authors: A
authors: A
authors: Alonso
authors: D
authors: D
authors: Rodríguez
authors: J M
authors: JM
authors: Morrell
authors: M
authors: M
authors: Miragaya
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002161:Camelids, New World / Q000196:embryology*; D002469:Cell Separation / Q000379:methods / Q000662:veterinary; D003470:Culture Media; D046149:Embryo Culture Techniques / Q000662:veterinary*; D047108:Embryonic Development / Q000502:physiology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D008297:Male; D009865:Oocytes / Q000502:physiology*; D012661:Semen / Q000166:cytology / Q000502:physiology; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology*; D020858:Tissue and Organ Harvesting / Q000379:methods / Q000662:veterinary
chemical_list: D003470:Culture Media