International Mammalian Genome Society

logo18th International Mouse Genome Conference

17-22 October 2004, Seattle, USA


ORAL PRESENTATION

THURSDAY OCTOBER 21

9.45am – 10.00am

RISK ASSESSMENT OF MOUSE HEPATITIS INFECTION (MHV) VIA IN VITRO FERTILIZATION AND EMBRYO TRANSFER WITH INTACT AND LASER MICRODISSECTED OOCYTES.

Peters D1, Marschall S1, Mahabir E2, Schmidt J2, Hrabé de Angelis M1

1 Institute of Experimental Genetics, GSF-National Research Center for Environmental and Health, Neuherberg, Germany, 2 Department of Comparative Medicine, GSF-National Research Center for Environmental and Health, Neuherberg, Germany

Cryobanking of mouse mutants and allocation of specific pathogen free (SPF) animals is of particular importance for the scientific community. Routinely used rederivation methods have to be enhanced by new assisted reproductive technologies (e.g. Zona dissection, ICSI) but these methods need to be investigated with regard to possible pathogen transmission.

The most common viral pathogen in mouse facilities, the Mouse Hepatitis Virus (MHV), was chosen to ensure a well-defined washing procedure used for in-vitro fertilization (IVF) and to estimate the risk increase by zona microdissection with PALM microbeam laser. Zona pellucida (ZP) intact and laser dissected oocytes used for the IVF were scheduled in three experiment groups:

oocytes in IVF were exposed to virus during fertilization and culture without washing procedure

oocytes in IVF were exposed to virus during fertilization and culture with washing procedure

laser microdissected oocytes in IVF were exposed to virus during fertilization and culture with washing procedure

Resulting embryos were transferred to SPF recipients, sera from recipients and offspring were tested for MHV-antibodies using ELISA. All used media were tested for presence and infectivity of MHV.

In experiment 1 nearly all recipients showed immune response whereas in experiment 2 and 3 neither in recipients nor in offspring antibodies against MHV could be detected.

Appropriate washing procedures applied in IVF protect against transmission of MHV even if the zona pellucida – the shield of the embryo – is disrupted.

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