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近交系BALB/cJ小鼠与远交系NMRI小鼠的超排卵、体外受精(IVF)和体外发育(IVD)方案比较

Superovulation, in vitro fertilization (IVF) and in vitro development (IVD) protocols for inbred BALB/cJ mice in comparison with outbred NMRI mice.

作者信息

Golkar-Narenji Afsaneh, Gourabi Hamid, Eimani Hussein, Barekati Zeinab, Akhlaghi Aliasghar

机构信息

Department of Genetics Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR No 12, Hafez St, Banihashem St, Resalat Ave 19395-4644 Tehran Iran.

Department of Embryology Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR Tehran Iran.

出版信息

Reprod Med Biol. 2012 Apr 7;11(4):185-192. doi: 10.1007/s12522-012-0127-8. eCollection 2012 Oct.

Abstract

PURPOSE

To study assisted reproductive technology (ART) protocols including superovulation, in vitro fertilization (IVF) and in vitro development (IVD) for BALB/cJ mice in comparison with a common ART protocol for NMRI mice.

METHODS

Adult NMRI and BALB/cJ mice were superovulated using a 48 h G-interval. In order to find a more suitable G-interval for the BALB/cJ strain, G-intervals including 44, 46 and 50 h were also examined. Superovulation rates were recorded in all groups. IVF rate of BALB/c oocytes in T6 and mHTF media were compared. IVD rates of BALB/cJ zygotes in mHTF, T6 and G1V/G2V media were compared. In addition, IVF and IVD rates of BALB/cJ and NMRI oocytes were compared in T6 medium during IVF-IVD procedures.

RESULTS

In BALB/cJ mice the highest superovulation rates were observed with 44-46 h G-intervals. However, with a 48 h G-interval, superovulation rates were significantly lower in BALB/cJ compared to NMRI mice ( < 0.05). mHTF medium significantly increased in vitro fertilization of BALB/cJ oocytes compared to T6 medium ( < 0.05). Fertilization rate of NMRI oocytes was significantly higher than BALB/cJ oocytes in T6 medium ( < 0.05). The BALB/cJ embryo IVD was significantly higher in G1/G2 medium compared to mHTF and T6 media ( < 0.01).

CONCLUSIONS

Superovulation with 48 h G-interval and using T6 during all in vitro procedures produces embryos more efficiently for NMRI mice than for BALB/cJ mice. For BALB/cJ mice, a protocol including superovulation with a 44-46 h G-interval, using mHTF during IVF and G1V/G2V medium during IVD, may improve in vitro embryo production.

摘要

目的

研究用于BALB/cJ小鼠的辅助生殖技术(ART)方案,包括超排卵、体外受精(IVF)和体外发育(IVD),并与用于NMRI小鼠的常见ART方案进行比较。

方法

成年NMRI和BALB/cJ小鼠采用48小时的促性腺激素间隔进行超排卵。为了找到更适合BALB/cJ品系的促性腺激素间隔,还检测了包括44、46和50小时的促性腺激素间隔。记录所有组的超排卵率。比较BALB/c卵母细胞在T6和mHTF培养基中的体外受精率。比较BALB/cJ受精卵在mHTF、T6和G1V/G2V培养基中的体外发育率。此外,在体外受精-体外发育过程中,比较BALB/cJ和NMRI卵母细胞在T6培养基中的体外受精率和体外发育率。

结果

在BALB/cJ小鼠中,促性腺激素间隔为44 - 46小时时观察到最高的超排卵率。然而,促性腺激素间隔为48小时时,BALB/cJ小鼠的超排卵率显著低于NMRI小鼠(<0.05)。与T6培养基相比,mHTF培养基显著提高了BALB/cJ卵母细胞的体外受精率(<0.05)。在T6培养基中,NMRI卵母细胞的受精率显著高于BALB/cJ卵母细胞(<0.05)。与mHTF和T6培养基相比,BALB/cJ胚胎在G1/G2培养基中的体外发育率显著更高(<0.01)。

结论

对于NMRI小鼠,促性腺激素间隔为48小时并在所有体外操作中使用T6比对于BALB/cJ小鼠能更有效地产生胚胎。对于BALB/cJ小鼠,一种包括促性腺激素间隔为44 - 46小时进行超排卵、体外受精时使用mHTF以及体外发育时使用G1V/G2V培养基的方案,可能会提高体外胚胎产量。

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