Comparison of two co-culture systems to assess the survival of in vitro produced bovine blastocysts after vitrification
(1998) Animal Reproduction Science — Vol. 52, n° 1, p. 39-50 (1998)
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- The ability of bovine blastocysts to recover after cryopreservation and thawing procedures is often assessed by evaluating their re-expansion during in vitro co-culture. However, the influence of factors such as feeder cell type and gas atmosphere on blastocyst survival and evolution have never been considered. This study therefore compared two cell co-culture systems and two different gas atmospheres to assess survival of in vitro produced bovine blastocysts after vitrification. Day-7 blastocysts (n = 181) were vitrified in a mixture of 25% glycerol/25% ethylene glycol. After warming and dilution, they were co-cultured either on Buffalo rat liver cells (BRL CC cell line) or on granulosa cells (GR CC primary culture) in TCM 199 supplemented with 10% FCS and under an atmosphere of 5% or 20% O-2. Surviving and hatching rates were recorded at 24 h intervals for 3 days. After 72 h of culture, surviving blastocysts were treated for differential counting of inner cell mass (ICM) and trophectoderm cells. Blastocyst survival rates were higher when BRL and granulosa co-culture were performed under 20% oxygen as compared to 5% oxygen (20% O-2: 62% vs. 5% O-2: 25%, P < 0.0001). However, the quality of blastocysts surviving in the granulosa co-culture condition was lower under 20% O-2 than under 5% O-2 as indicated by lower total and trophectoderm cell numbers (respectively 79 +/- 6 and 56 +/- 6 at 20% O-2 vs. 100 +/- 10 and 74 +/- 10 at 5% O-2, P < 0.05), by an altered ICM/trophectoderm ratio (20% O-2: 28% vs. 5% O-2: 23%, P < 0.05), by a higher total nuclear fragmentation (20% O-2: 13.7% vs. 5% O-2: 1.5%. P < 0.05) and a trend to decreased hatching (20% O-2: 32% vs. 5% O-2: 81%, P = 0.07). Whereas, for BRL co-culture, 20% O-2 yielded higher quality blastocysts than 5% O-2 as evaluated by higher ICM and trophectoderm cell numbers (19 +/- 1 and 71 +/- 5 at 20% O-2 vs. 15 +/- 2 and 48 +/- 9 at 5% O-2, respectively, P < 0.05), by lower nuclear fragmentation in the ICM (20% O-2: 2.2% vs. 5% O-2: 6.7%, P < 0.05). In conclusion, co-culture conditions may influence blastocysts survival and quality after cryopreservation. In our conditions, co-culture with BRL cells under 20% O-2 seems to be the best combination to evaluate blastocyst survival and quality after vitrification. (C) 1998 Elsevier Science B.V. All rights reserved.
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Kaidi, S., Donnay, I., Van Langendonckt, A., Dessy, F., & Massip, A. (1998). Comparison of two co-culture systems to assess the survival of in vitro produced bovine blastocysts after vitrification. Animal Reproduction Science, 52(1), 39-50. https://doi.org/10.1016/S0378-4320(98)00089-X (Original work published 1998)