Sperm And Embryo Cryopreservation
Sperm cryopreservation
This is the fastest and least expensive option for cryopreservation of mouse strains. We can store frozen sperm in LN2 indefinitely. We can generally reconstitute live mice using IVF. Our team freezes and stores 10 samples in liquid nitrogen following collection of epididymal sperm. We will keep them in two separate locations.
Embryo cryopreservation
This procedure is ideal for preserving multiple homozygous traits (>3). Currently, fewer than 1% of researchers at OHSU use this service.
- IVF embryo cryopreservation (Speed Cryo): Our team cryopreserves the freshly collected sperm from 2-4 transgenic males as with sperm cryopreservation. We then use this sperm to fertilize the eggs of super-ovulated 3–4-week-old females of the same genotype via in vitro fertilization and then cryopreservation of 2 cell or 6-8 cell stage embryos is performed. Typically, you can expect 10 frozen embryos per female provided for this procedure. It then takes 25-30 embryos to resuscitate a line. This method is generally only recommended when preserving multiple homozygous genes is desired.
- Standard embryo cryopreservation: For strain cryopreservation, 10-12 homozygous or heterozygous stud males will mate with super-ovulated wildtype females. Our team will collect approximately 250-300 8-cell stage embryos and freeze them. We will store roughly 20-30 embryos per straw. We can then store them in liquid nitrogen in the Core’s freezers. Freezing of 8-cell embryos is the most stable stage for embryo cryopreservation and storage.
Quality control
We can check each cryopreserved strain to ensure the overall quality and viability of embryos or sperm using one of three methods:
- After sperm cryopreservation, our team will thaw one straw and visually evaluate the quality of the spermatozoa under a microscope.
- Following standard or Speed Cryo freezing, our team will thaw one straw and place the embryos in a culture for development to blastocysts. A >75% development to blastocysts indicates a successful cryopreservation.
- Following standard or Speed Cryo freezing, our team will thaw one straw, and place the embryos in a culture for development to blastocysts followed by completing a uterine transfer into a pseudo-pregnant recipient. The generation of live pups displaying the appropriate genotype (genotyping to be done by the customer) indicates a successful cryopreservation.
Cryostorage
The Animal Model Support Core (AMSC) can store investigator-provided or AMSC-harvested embryos or sperm in liquid nitrogen in two freezers located at two geographically distinct sites. One site is at the OHSU Main Campus LBRB building and the other site is at the OHSU Main Campus RJH building.
Cost
- Cryopreservation for embryos
Our team freezes 20-30 eight-cell stage embryos per two female mice provided.
$1870
Speed cryo cryopreservation service fees include:
- Cryopreservation of two cell embryos based on number of females provided.
- Cryopreservation of 10 samples of sperm.
- Cryostorage of frozen embryos and sperm for 1 month.
Standard embryo cryopreservation service fees include
- Purchase of 3-week-old females (donors of oocytes).
- Cryopreservation of 250-300 eight-cell embryos.
- Cryostorage of frozen embryos for 1 month.
- Quality control for frozen embryos
Our team generates live 14-day old fetuses by retransfer of thawed embryos into the uterus of a foster dam.
$300
- Cryopreservation for sperm
Our team freezes 10 vials of sperm. Service fee includes cryostorage of frozen sperm 1 month.
$700
- Quality control for frozen sperm
Our team will thaw one vial and evaluate sperm quality visually.
$50
- Cryostorage for embryos, sperm or embryonic stem cells (lines 1 to 10)
Our team will cryostore 10-20 vials or straws, 1 line per year
$120
- Cryostorage for embryos, sperm or embryonic stem cells (lines 11 to 20)
Our team will cryostore 10 vials or straws, 1 line per year
$90
- Cryostorage for embryos, sperm or embryonic stem cells (lines 21 to 30)
Our team will cryostore 10 vials or straws, 1 line per year
$60
- Cryostorage for embryos, sperm or embryonic stem cells (lines 31 to 50)
Our team will cryostore 10 vials or straws, 1 line per year
$30
- Cryostorage for embryos, sperm or embryonic stem cells (lines 51 or more)
Our team will cryostore 10 vials or straws, 1 line per year
$12
Standard prices may vary depending on mouse donor strain and client’s affiliation. Changes will be based on the cost of additional reagents, mice and time of personnel of ASMC. External academic clients and commercial clients must pay an extra fee of 15% and 50% respectively.
Requesting services
Go to the AMSC iLab to request services. Please provide the following materials in your request:
- For Sperm Cryopreservation, provide 2-4 stud males at the age of 3-9 months.
- For Speed Cryo, provide a minimum of 6 breeding age males as well as 6 females age 3-4 weeks.
- For standard cryopreservation of 8-cell stage embryos, provide 10-12 stud males 2-9 months old.