Download CTC Embryo Cryopreservation Request Form

CBS Transgenic Core (CTC)
APPLICATION FOR CTC CRYOPRESERVATION OF EMBRYOS
Important information

Please complete one form for each line to be frozen.

Preparation for embryo cryopreservation is more time consuming than for sperm freezing
and it may require several sessions depending on the method used. However, the
rederivation process is much easier (just requiring surgical transfer into recipient females).

The procedure is to freeze compacted 8 Cells/Morula Stage Embryos using the Vitrification
method (Media - Ethylene Glycol/Ficoll + 0.5M Sucrose) in straws.

The number of eggs recommended will depend on the genotypes of your line (i.e.
homozygotes would require less than heterozygotes). We aim to freeze ~200 embryos for
each line.

We assume 35%> 55% of transferred blastocysts will go to live offspring

Embryo stocks can be replenished after re-establishing a line in-house if requested by the
researcher.

The researcher is responsible for providing appropriately aged males and females.

Female mice should be aged between 3-6 weeks in order to obtain the optimal embryo
yields

There are 2 options for embryo cryopreservation:
a)
A cohort of 20 (with a minimum of 10) females and mated with and a cohort of
singly housed stud males and the resulting embryos are collected and frozen. This
may require several sessions with new females in order to achieve a suitable number
of embryos to be frozen. The cost for this is based on the number of cryopreservation
sessions required.
b)
A cohort of 3-6 females (age 3-4 weeks) are superovulated and eggs collected. A
male mouse is sacrificed and sperm collected for an IVF procedure. All eggs are
frozen. This is approximately the same cost as a normal IVF session

CTC will store embryo straws until the researcher leaves the UoE, at which time, they must
make inform the CTC manager what should be done with them.

For quality control, wild type embryos are test frozen, thawed and cultured to blastocyst to
test each batch of embryo media before transgenic embryos are frozen.
To be completed and returned to [email protected]
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CBS Transgenic Core (CTC)
SECTION A:
Group Leader name and email:
Lab contact name and email:
Date of application:
PPL NUMBER:
Transgenic request number (if rederived or
generated by CTC e.g.TR-20-07):
SECTION B:
Name of female line to be used (i.e. your
transgenic line or wildtype females. If wild type please
supply the exact genetic background required)
Name of male line to be used (i.e. your transgenic
line or wildtype male. If wild type please supply the exact
genetic background required)
Type of service: Natural matings with collection of
eggs over several sessions or IVF
SECTION C:
Location of Animals
Tick@lab line name
Tick@lab team name
Nature of genetic modification and details:
(e.g. Transgenic, Knock out/in, spontaneous mutant,
induced mutant, etc with details of how was the stock
produced: Pronuclear injection/homologous
recombination/ gene trapping/other.) Please give details of
all alleles carried by the line if there is a combination.
Genetic Background: (C57Bl6/CBA/F1 hybrid etc).
No. backcrosses (if applicable):
Phenotype:
Expected genotypes: Please give details of all the
expected genotypes and predicted ratios on thawing if
more than one allele is expected
Reproductive performance (is the line difficult to
breed? Are homozygous mice viable/fertile?)
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CBS Transgenic Core (CTC)
Special husbandry requirements (if applicable):
Expression pattern of transgene (if known):
Coat Colour:
PCR primer sequences used for genotyping:
(Also specify reagents, conditions, thermocycling
parameters)
Please give details of size and sequence of
PCR product:
Description of genotyping method if other
than PCR:
References:
To be completed by CTC staff:
CTC Cryopreservation number:
CTC Technician who performed freezing and QC:
Location of long term storage: IGMM/LF (delete as appropriate)
QUALITY CONTROL MEDIA THAW/CULTURE:
Date
Media
Batch
QC
Strain
Morula
Recovered
Viable /
%
Cultured
to Blasts
% to
Blast
Pass
Y/N
QUALITY CONTROL GENETIC STRAIN
Date
Media
Batch
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Strain
Morula
Recovered
Viable / %
Cultured
to Blasts
% to Blast