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AI InsightDevelopmental evaluations of in vitro cultured mouse embryos could be confusing when only the native route was employed, and a staining technique could give more reliable results.
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Özet
Various BSA (bovine serum albumin) levels and media effects on development of 2-cell stage mouse embryos in in vitro culture were investigated in this 2 stage study. In the first stage, 1400 2-celled mouse embryos were cultured in 2 various BSA-bearing culture media for 96 h and directly observed microscopically. In the second stage, 1024 2-celled mouse embryos cultured for the same period and cell numbers (nuclei of cells) were determined by staining techniques. Embryos were cultured in both stages in M 16 and Whitten's media containing 0 (control), 0.3, 1, 3, 9, 18 and 36 mg/ml of BSA. The highest blastocyst rate was 94.57% ± 7.43, observed in 3 mg/ml BSA-containing Whitten's medium group (P < 0.05). The highest hatching + hatched blastocyst rates were obtained in the same group with 67.70% ± 25.22. The highest nucleus number in the M 16 medium was in the 1 mg/ml BSA group (95.45 ± 9.61). In the M16 medium group, the 3 mg/ml BSA group showed the highest development rate in the first stage of the study; however, the nucleus numbers in this group (81.37 ± 18.31), were significantly lower than the nucleus numbers of the 1 mg/ml BSA group (P < 0.05). It was concluded that the optimal benefit to in vitro culture of 2-cell mouse embryos could be maintained when BSA was used as a protein source at 1 mg/ml and 3 mg/ml doses, and that M16 or Whitten's media could be used successfully as a culture medium. Developmental evaluations of in vitro cultured mouse embryos could be confusing when only the native route was employed, and a staining technique could give more reliable results.
Abstract
Various BSA (bovine serum albumin) levels and media effects on development of 2-cell stage mouse embryos in in vitro culture were investigated in this 2 stage study. In the first stage, 1400 2-celled mouse embryos were cultured in 2 various BSA-bearing culture media for 96 h and directly observed microscopically. In the second stage, 1024 2-celled mouse embryos cultured for the same period and cell numbers (nuclei of cells) were determined by staining techniques. Embryos were cultured in both stages in M 16 and Whitten's media containing 0 (control), 0.3, 1, 3, 9, 18 and 36 mg/ml of BSA. The highest blastocyst rate was 94.57% ± 7.43, observed in 3 mg/ml BSA-containing Whitten's medium group (P < 0.05). The highest hatching + hatched blastocyst rates were obtained in the same group with 67.70% ± 25.22. The highest nucleus number in the M 16 medium was in the 1 mg/ml BSA group (95.45 ± 9.61). In the M16 medium group, the 3 mg/ml BSA group showed the highest development rate in the first stage of the study; however, the nucleus numbers in this group (81.37 ± 18.31), were significantly lower than the nucleus numbers of the 1 mg/ml BSA group (P < 0.05). It was concluded that the optimal benefit to in vitro culture of 2-cell mouse embryos could be maintained when BSA was used as a protein source at 1 mg/ml and 3 mg/ml doses, and that M16 or Whitten's media could be used successfully as a culture medium. Developmental evaluations of in vitro cultured mouse embryos could be confusing when only the native route was employed, and a staining technique could give more reliable results.