Short-term preservation of canine preantral follicles: Effects of temperature, medium and time.
The use of the large pool of preantral follicles is a promising alternative to provide high numbers of fertilizable oocytes to reproductive biotechnology. This issue is particularly important to canids, since current rates of success of in vitro techniques using oocytes are very limited, and many species within this family are threatened by extinction. The aim of this study was to evaluate effects of temperature, medium and time on morphology and viability of canine preantral follicles during short-term preservation. Canine ovaries were cut into fragments which were incubated in 0.9% NaCl solution or in minimum essential medium (MEM) at 4, 20 or 38 degrees C for 2, 6, 12 or 24 h. Afterwards, preantral follicles were analyzed by histology, transmission electron microscopy and viability testing using trypan blue, calcein-AM and ethidium homodimer-1. Percentages of morphological normal and viable follicles were maintained similar to control (time 0 h) after incubation in 0.9% NaCl at 4 or 20 degrees C for up to 6h and at 38 degrees C for 2 h. Using MEM, such preservation was possible for 12h at 4 or 20 degrees C, and for 6h at 38 degrees C. These results indicate that preservation of canine preantral follicles might be better accomplished through hypothermic (4 or 20 degrees C) storage in MEM, which ensures maintenance of morphology and viability for up to 12h.
Lopes
Cláudio Afonso Pinho
CA
Faculty of Veterinary, PPGCV, LAMOFOPA, State University of Ceará, Av. Paranjana 1700, Campus do Itaperi, CEP 60.740-903, Fortaleza, CE, Brazil. lopes@izw-berlin.de
dos Santos
Regiane Rodrigues
RR
Celestino
Juliana Jales de Hollanda
JJ
Melo
Mônica Aline Parente
MA
Chaves
Roberta Nogueira
RN
Campello
Claudio Cabral
CC
Silva
José Roberto Viana
JR
Báo
Sônia Nair
SN
Jewgenow
Katarina
K
de Figueiredo
José Ricardo
JR
D000818:Animals; D002470:Cell Survival; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D006107:Granulosa Cells / Q000648:ultrastructure; D008928:Mitochondria / Q000648:ultrastructure; D009685:Nuclear Envelope / Q000648:ultrastructure; D009865:Oocytes / Q000502:physiology*; D009926:Organ Preservation / Q000379:methods / Q000662:veterinary; D006080:Ovarian Follicle / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D010052:Ovariectomy / Q000662:veterinary