Karja N W Kurniani, Otoi Takeshige, Murakami Masako, Yuge Minori, Fahrudin Mokhamad, Suzuki Tatsuyuki
Department of Veterinary Sciences, Yamaguchi University, Japan.
Reprod Fertil Dev. 2002;14(5-6):291-6. doi: 10.1071/rd01135.
The effects of protein supplementation in culture medium on development to the hatching and hatched blastocyst stages of cat in vitro-fertilized embryos were investigated. In the first experiment, presumptive zygotes derived from in vitro maturation and in vitro fertilization (IVF) were cultured in modified Earle's balanced salt solution (MK-1) supplemented with 0.4% bovine serum albumin (BSA) or 5% fetal bovine serum (FBS) for 9 days. There were no significant differences between the BSA and FBS groups with respect to the proportion of cleavage and development to the morula and blastocyst stages of zygotes. However, the presence of FBS in the medium enhanced development to the hatching blastocyst stage of zygotes compared with the BSA group (31.4% v. 7.8%). Moreover, 2.9% of zygotes cultured with FBS developed to the hatched blastocyst stage. The mean cell number of blastocysts derived from zygotes cultured with FBS was significantly higher (P<0.01) than that from zygotes cultured with BSA (136.6 v.101.5). In the second experiment, embryos at the morula orblastocyst stage, which were produced by culturing in MK-1 supplemented with 0.4% BSA after IVF, were subsequently cultured in MK-1 with 0.4% BSA or 5% FBS. Significantly more morulae developed to the blastocyst (P<0.05) and hatching blastocyst stages (P<0.01) in the FBS group than in the BSA group (71.5% and 53.6% v. 44.9% and 6.0%, respectively). Although none of the morulae cultured with BSA developed to the hatched blastocyst stage, 11.5% of morulae cultured with FBS developed to the hatched blastocyst stage. Moreover, the proportion of development to the hatching blastocyst stage of blastocysts was significantly higher (P<0.01) in the FBS group than in the BSA group (68.7% v. 9.8%). None of the blastocysts cultured with BSA developed to the hatched blastocyst stage, whereas 7.3% of blastocysts cultured with FBS developed to the hatched blastocyst stage. The results of the present study indicate that supplementation with FBS at different stages of early embryo development promotes development to the hatching and hatched blastocyst stages of cat IVF embryos.
研究了在培养基中补充蛋白质对猫体外受精胚胎发育至孵化和孵化后囊胚阶段的影响。在第一个实验中，将体外成熟和体外受精（IVF）获得的假定受精卵在添加0.4%牛血清白蛋白（BSA）或5%胎牛血清（FBS）的改良Earle平衡盐溶液（MK-1）中培养9天。在受精卵的卵裂比例以及发育至桑葚胚和囊胚阶段方面，BSA组和FBS组之间没有显著差异。然而，与BSA组相比，培养基中添加FBS可提高受精卵发育至孵化囊胚阶段的比例（31.4%对7.8%）。此外，用FBS培养的受精卵中有2.9%发育至孵化囊胚阶段。用FBS培养的受精卵来源的囊胚平均细胞数显著高于（P<0.01）用BSA培养的受精卵来源的囊胚（136.6对101.5）。在第二个实验中，将IVF后在添加0.4%BSA的MK-1中培养产生的桑葚胚或囊胚阶段的胚胎，随后在添加0.4%BSA或5%FBS的MK-1中培养。FBS组中发育至囊胚（P<0.05）和孵化囊胚阶段（P<0.01）的桑葚胚显著多于BSA组（分别为71.5%和53.6%对44.9%和6.0%）。虽然用BSA培养的桑葚胚均未发育至孵化囊胚阶段，但用FBS培养的桑葚胚中有11.5%发育至孵化囊胚阶段。此外，FBS组中囊胚发育至孵化囊胚阶段的比例显著高于（P<0.01）BSA组（68.7%对9.8%）。用BSA培养的囊胚均未发育至孵化囊胚阶段，而用FBS培养的囊胚中有7.3%发育至孵化囊胚阶段。本研究结果表明，在早期胚胎发育的不同阶段补充FBS可促进猫IVF胚胎发育至孵化和孵化后囊胚阶段。
