OPU-IVF PROTOCOLS

SAR IVM

Oocytes Aspiration Plus

SAR IVF

SAR IVC

 

IVF for Large Scale Embryo Production PROTOCOLS

SAR IVM

SAR IVF

SAR IVC

 

 

 

 

 

 

 

 

 

 

OPU-IVF PROTOCOL

 

 

SAR IVM

 

Expiration Date: If continuously stored at 4C, two weeks after production date; If frozen upon arrival, 2-3 months at -20C.
Renova Life, Inc. (RLI) recommends that SAR-IVM maturation be stored at -20C upon arrival, this makes FSH/LH function as effectively as freshly prepared lots.

1.At the time of arrival from Renova, pipet 1 mL SAR-IVM into 1.8 mL cryovials and overlay with 100 L of oil into 1ml vials and place immediately into -20C freezer.

2. Remove one vial SAR IVM from freezer and place into 5% CO2 incubator at 39 C with the lid loose for equilibration overnight. One vial SAR IVM is for maturation of up to 25 or more oocytes per cow.

3.In the morning of OPU, remove vial(s) from incubator, screw cap on tight before removing from incubator and parafilm around cap, place immediately into 39 C warmed transport incubator.

4.Maintain vials at 39 C at all times.

 

Addendum:
*If oocytes will be aspirated and transported in under 2 h, oocytes can be stored in 1 mL vials of OWP (oocyte washing prior to IVM) media and held at 39 C in transport incubator. At the lab each cows oocytes should be washed through one 50 L drop of SAR-IVM before placing into SAR-IVM drop for maturation. For example: 1 cow = 1 (50 L) drop for washing + 1 (50 L) drop for maturation culture.

*If oocytes will be aspirated and transported in less than 6 h, oocytes can be transferred into SAR IVM pre-equilibrated media dishes (25 oocytes per 50uL drop). Dishes need to be pre-equilibrated for 2-3 hours at 5% CO2, 39 C.

* SAR IVM vials can be used within 48 h of CO2 equilibration. If this is the case, vials should be equilibrated overnight in CO2 incubator. Caps should be sealed and parafilmed and placed on ice. Warm vials 2 h before using.

 

Preparation at the clinic prior to OPU:

    • 1.Water bath is turned on and temperature is stabilized at 39 C. OAP and OWP media are placed in water bath and media temperature is brought up to 39 C prior to the beginning of OPU. Square grid dishes, 35mm plates, transfer pipets, filters, and hemostats are set-up near the stereomicroscope.

     

     

     

    OPU-IVF Protocol (pdf format)