Department of Veterinary Medicine, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan.
Cryobiology. 2011 Dec;63(3):170-4. doi: 10.1016/j.cryobiol.2011.06.002. Epub 2011 Aug 24.
The objectives of this study were to examine the effects of the presence or absence of serum during the in vitro culturing period of domestic cat embryos on their developmental potential into blastocysts as well as their tolerance to cryopreservation using a slow-freezing method. In vitro-fertilized cat oocytes were incubated in a modified synthetic oviduct fluid (mSOF) containing 4 mg/mL bovine serum albumin (BSA) throughout culturing (BSA group) or in mSOF containing 4 mg/mL BSA for the first 3 days followed by mSOF containing 5% fetal bovine serum (FBS group). The developmental potential of the embryos to the blastocyst and expanded blastocyst stages was evaluated 7 days after in vitro fertilization. The blastocysts were frozen-thawed by the slow-freezing method and cultured for 3 days to examine their viability in vitro. There were no differences in the formation rates of blastocysts or expanded blastocysts, or number of cells in the embryos between the two groups. After cryopreservation, the hatching rates of the expanded blastocysts in the BSA group were significantly higher (P<0.05) than those of the FBS group. The postthaw viability of blastocysts was lower than that of expanded blastocysts irrespective of culture medium. These results indicate that the developmental potential of cat embryos cultured in serum-free medium is comparable to those cultured in serum-containing medium. Furthermore, expanded blastocysts produced without serum exhibit better postthaw viability than those produced with serum.
本研究旨在探讨在体外培养家猫胚胎期间是否添加血清对其发育为囊胚的潜力以及对慢速冻法冷冻保存的耐受性的影响。体外受精的猫卵母细胞在含有 4 mg/mL 牛血清白蛋白(BSA)的改良合成输卵管液(mSOF)中培养,整个培养过程中添加 BSA(BSA 组)或前 3 天添加 mSOF 中添加 4 mg/mL BSA,然后添加 5%胎牛血清(FBS 组)。在体外受精后 7 天评估胚胎发育为囊胚和扩张囊胚阶段的潜力。通过慢速冻法冷冻和解冻囊胚,并培养 3 天,以检测其体外活力。两组之间的囊胚或扩张囊胚形成率或胚胎细胞数量均无差异。冷冻保存后,BSA 组扩张囊胚的孵化率显著高于 FBS 组(P<0.05)。无论培养基如何,解冻后囊胚的活力均低于扩张囊胚。这些结果表明,在无血清培养基中培养的猫胚胎的发育潜力与在含血清培养基中培养的胚胎相当。此外,无血清产生的扩张囊胚的解冻后活力优于含血清产生的扩张囊胚。