A new selection criterion to assess good quality ovine blastocysts after vitrification and to predict their transfer into recipients

Molecular Reproduction and Development - Tập 75 Số 2 - Trang 373-382 - 2008
Giovanni Giuseppe Leoni1, Fiammetta Berlinguer2, Sara Succu2, Daniela Bebbere2, F. Mossa2, Manuela Madeddu2, S. Ledda3, Luisa Bogliolo3, Salvatore Naitana2
1Department of Physiological, Biochemical and Cellular Sciences, University of Sassari, Sassari, Italy
2Dept. of Animal Biology, University of Sassari, Sassari, Italy
3Institute of General Pathology, Pathological Anatomy and Obstetric-Surgery Veterinary Clinic, University of Sassari, Sassari, Italy

Tóm tắt

AbstractThe feasibility to accurately select viable embryos would be valuable for improving pregnancy rates and avoiding futile transfer attempts. The aim of our study was to assess if in vitro‐produced embryo quality could be determined by the timing of blastocoelic cavity re‐expansion after vitrification, warming, and in vitro culture using sheep as a model. Blastocysts were produced in vitro, vitrified/warmed, and cultured in TCM‐199 plus 10% FCS for 72 hr. Embryos were divided into two groups: re‐expanded within 8 hr (A) and from 8 to 16 hr (B) of IVC after warming. Fast re‐expanded blastocysts showed higher in vitro hatching rates and total cell number calculated on the hatched blastocysts compared with slow re‐expanded ones (P < 0.01). Peroxide status evaluation (P < 0.01) and TUNEL test (P < 0.05) revealed a higher number of positive cells in group B compared with group A. The quantitative analysis of protein synthesis revealed a higher synthesis in fast compared with slow re‐expanded embryos (P < 0.05). Quantitative RT‐PCR showed that 90‐kDa Heat Shock Protein β was more expressed in group A than in group B (P < 0.05), while the quantity of P34cdc2, Cyclin b, Aquaporin 3, Na/K ATPase, and Actin did not differ between the two groups. Pregnancy rates after transfer to synchronized recipients were higher in fast compared to slow re‐expanded blastocysts (P < 0.05). Our results evidenced that timing of blastocoelic cavity re‐expansion after vitrification/warming and in vitro culture can be considered as a reliable index of in vitro produced embryo quality and developmental potential. Mol. Reprod. Dev. 75: 373–382, 2008. © 2007 Wiley‐Liss, Inc.

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Tài liệu tham khảo

Arav A, 1992, New trends in embryo transfer, 255

10.1002/mrd.10388

10.1002/mrd.20263

10.1016/S0012-1606(02)00127-6

10.1006/dbio.1998.8874

10.1002/mrd.20450

10.1016/0167-5699(94)90018-3

10.1093/humrep/17.9.2419

10.1242/dev.121.1.113

10.1016/S0093-691X(02)01270-0

10.1016/S1472-6483(10)60842-5

10.1126/science.8100364

10.1016/0093-691X(95)00351-8

10.1051/rnd:19990501

10.1095/biolreprod.105.045823

10.1016/S0960-9822(01)00329-3

10.1016/j.cryobiol.2005.08.002

10.1093/ilar.41.4.187

10.1002/(SICI)1098-2795(199608)44:4<472::AID-MRD6>3.0.CO;2-I

10.1016/S0015-0282(01)02888-6

Halliwell B, 1989, Free radicals in biology and medicine, 22

10.1073/pnas.98.4.1655

10.1002/1098-2795(200012)57:4<353::AID-MRD7>3.0.CO;2-R

10.1016/0093-691X(94)00020-U

10.1002/mrd.1080370305

10.1016/S0378-4320(98)00089-X

10.1095/biolreprod65.4.1127

10.1016/S1472-6483(10)62125-6

10.1093/humrep/17.7.1863

10.1006/cryo.2001.2335

10.1038/227680a0

10.1093/humrep/17.10.2686

10.1530/rep.1.00878

10.1095/biolreprod.102.004481

10.1016/S0011-2240(02)00139-6

10.1002/mrd.10251

10.1016/j.anireprosci.2005.05.027

10.1073/pnas.79.10.3218

10.1095/biolreprod.102.006833

Lindquist S, 1984, Heat shock—A comparison of Drosophila and yeast, J Embryol Exp Morphol, 83, 147

Liu AY, 1994, Transient cold shock induces the heat shock response upon recovery at 37°C in human cells, J Biol Chem, 269, 14768, 10.1016/S0021-9258(17)36691-7

10.1016/j.theriogenology.2005.09.028

10.1007/PL00000590

10.1016/S0378-4320(97)00043-2

10.1095/biolreprod.104.027987

10.1095/biolreprod.103.022152

10.1016/S0378-4320(02)00162-8

10.1017/S0967199406003649

10.1016/0011-2240(87)90042-3

10.1530/jrf.0.1010681

10.1002/mrd.10138

10.1095/biolreprod.102.007799

10.1002/mrd.20133

10.1095/biolreprod.102.006320

10.1093/humrep/deg184

Tervit HR, 1972, Successful culture in vitro of sheep and cattle ova, J Reprod Fertil, 30, 487

10.1016/S1472-6483(10)61091-7

10.1016/0093-691X(95)00414-4

10.1002/(SICI)1098-2795(199705)47:1<47::AID-MRD7>3.0.CO;2-Q

10.1095/biolreprod55.3.703

10.1002/dvg.1020140207

10.1007/s10254-003-0021-1

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Molecular Reproduction and Development

Tập 75 Số 2

373-382

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