Dataset Information

Conserving, distributing and managing genetically modified mouse lines by sperm cryopreservation.

ABSTRACT:

Background

Sperm from C57BL/6 mice are difficult to cryopreserve and recover. Yet, the majority of genetically modified (GM) lines are maintained on this genetic background.

Methodology/principal findings

Reported here is the development of an easily implemented method that consistently yields fertilization rates of 70+/-5% with this strain. This six-fold increase is achieved by collecting sperm from the vas deferens and epididymis into a cryoprotective medium of 18% raffinose (w/v), 3% skim milk (w/v) and 477 microM monothioglycerol. The sperm suspension is loaded into 0.25 mL French straws and cooled at 37+/-1 degrees C/min before being plunged and then stored in LN(2). Subsequent to storage, the sperm are warmed at 2,232+/-162 degrees C/min and incubated in in vitro fertilization media for an hour prior to the addition of oocyte cumulus masses from superovulated females. Sperm from 735 GM mouse lines on 12 common genetic backgrounds including C57BL/6J, BALB/cJ, 129S1/SvImJ, FVB/NJ and NOD/ShiLtJ were cryopreserved and recovered. C57BL/6J and BALB/cByJ fertilization rates, using frozen sperm, were slightly reduced compared to rates involving fresh sperm; fertilization rates using fresh or frozen sperm were equivalent in all other lines. Developmental capacity of embryos produced using cryopreserved sperm was equivalent, or superior to, cryopreserved IVF-derived embryos.

Conclusions/significance

Combined, these results demonstrate the broad applicability of our approach as an economical and efficient option for archiving and distributing mice.

SUBMITTER: Ostermeier GC

PROVIDER: S-EPMC2453316 | biostudies-literature | 2008 Jul

REPOSITORIES: biostudies-literature

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Publications

Conserving, distributing and managing genetically modified mouse lines by sperm cryopreservation.

Ostermeier G Charles GC Wiles Michael V MV Farley Jane S JS Taft Robert A RA

PloS one 20080730 7

<h4>Background</h4>Sperm from C57BL/6 mice are difficult to cryopreserve and recover. Yet, the majority of genetically modified (GM) lines are maintained on this genetic background.<h4>Methodology/principal findings</h4>Reported here is the development of an easily implemented method that consistently yields fertilization rates of 70+/-5% with this strain. This six-fold increase is achieved by collecting sperm from the vas deferens and epididymis into a cryoprotective medium of 18% raffinose (w/ ...[more]

PMID: 18665210

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Dataset Information

Conserving, distributing and managing genetically modified mouse lines by sperm cryopreservation.

Background

Methodology/principal findings

Conclusions/significance

Publications

Conserving, distributing and managing genetically modified mouse lines by sperm cryopreservation.

Similar Datasets

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