Superovulation, in vitro fertilization (IVF) and in vitro development (IVD) protocols for inbred BALB/cJ mice in compari

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ORIGINAL ARTICLE

Superovulation, in vitro fertilization (IVF) and in vitro development (IVD) protocols for inbred BALB/cJ mice in comparison with outbred NMRI mice Afsaneh Golkar-Narenji • Hamid Gourabi Hussein Eimani • Zeinab Barekati • Aliasghar Akhlaghi



Received: 28 November 2011 / Accepted: 25 March 2012 / Published online: 7 April 2012 Ó Japan Society for Reproductive Medicine 2012

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 G1V5/ G2V5 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 (p \ 0.05). mHTF medium significantly increased in vitro fertilization of BALB/cJ oocytes compared to T6 medium (p \ 0.05). Fertilization rate of NMRI oocytes was significantly higher than BALB/cJ oocytes in T6 medium (p \ 0.05). The BALB/cJ embryo IVD was significantly higher in G1/G2 medium compared to mHTF and T6 media (p \ 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 G1V5/G2V5 medium during IVD, may improve in vitro embryo production.

A. Golkar-Narenji  H. Gourabi (&) 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 e-mail: [email protected]

Keywords In vitro development (IVD)  In vitro fertilization (IVF)  Medium  Mouse strain  Superovulation

H. Eimani Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran Z. Barekati Laboratory for Gynecological Oncology, Department of Biomedicine, Women’s Hospital, University of Basel, Basel, Switzerland A. Akhlaghi Department of Epidemiology and Reproductive Health, Reproductive epidemiology Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

Introduction The inbred BALB/c mouse strain was established in 1935 [1] and is widely used for genetic research [2] and genetic engineering [3]. This strain has also been used in tumor and immunologic research. BALB/c is also