山羊卵母细胞无血清体外成熟培养研究

摘要：为了研究无血清培养系统对山羊卵母细胞体外成熟及孤雌激活胚胎早期发育力的影响，分别以0.3% BSA(m/v)、1% PVA(m/v)、1% ITS(v/v)代替成熟培养液中FBS，以添加5% FBS的OM为对照组，对来源于屠宰场的COCs进行体外培养。成熟率PVA添加组（50.00%），显著低于FBS组（83.23%）（P﹤0.01），BSA组（71.19%）和ITS组（76.79%）与FBS组无显著差异（P＞0.05）；成熟卵母细胞用离子霉素联合6-DMAP激活，在SOFaa中进行孤雌胚培养，BSA、PVA、ITS及 FBS各组的卵裂率依次为61.90%、46.97%、79.07%和83.58%，囊胚率分别为25.00%、19.70%、55.81%和60.45%。实验结果指出:在OM基础培养液中添加BSA或PVA卵母细胞的成熟率明显低于添加FBS，且卵裂率和囊胚发育率也低于对照组(P<0.05或 P<0.01)；添加ITS，卵母细胞成熟率虽低于对照组，但孤雌胚卵裂率和囊胚发育率与对照组接近（P>0.05）。结论：以ITS代替FBS，可以用于山羊卵母细胞体外成熟培养。

In Vitro Serum-free Maturation Culture of Goat Oocytes

Abstract: To evaluate the effects of a system of serum-free maturation culture on in vitro maturation of goat oocytes and development of parthenogenesis embryos，0.3 % BSA (m/v), 1% PVA (m/v) and 1% ITS (v/v) were added to basic medium of oocyte maturation as substitute of serum, 5% fetal bovine serum was added to control. Cumulus-oocyte complexes (COCS) derived abattoir ovaries were cultured in vitro with those culture medium respectively. Maturation rate of PVA supplementation group (50.00%) was significant lower than that of control (83.23%)(P<0.01), and that of BSA group (71.19%)and ITS group(76.79%) had no difference compared to control; The matured oocytes were actived by ionoycin and 6-dimethylaminopurine, then, parthenogenesis embryos were cultured in SOFaa medium to observe development of embryos each group. Cleavage rate from BSA, PVA, ITS and FBS group was 61.90%, 46.97%, 79.07% and 83.58% respectively, and blastocyst rate respectively was 25.00%, 19.70%, 55.81% and 60.45%. The results show that substitute PVA and BSA for FBS in elemental maturation medium decrease significantly the abilities of maturation and early development of embryos(P<0.05 or P<0.01),substitute ITS for FBS decrease the abilities of maturation ,but has no effect on early development of embryos(P>0.05).Conclusion: ITS may be applied to in vitro maturation of goat oocyte as substitute of FBS.