PREPARATION FOR IN VITRO PRODUCTION OF EMBRYOS
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Materials and Equipment Needed
Stock Solutions
Media Preparation
2 x 1 L sterile saline
(transport saline)
1
aliquot of stock 16 (Pen-Strep 10 ml) per bottle of saline
1 bottle of oocyte collection
medium (OCM) (400 ml)
1
aliquot of stock 11 (glutamine 4 ml) per bottle of OCM
1
aliquot of stock 15 (Pen/Strep 4 ml) per bottle of OCM
1
aliquot of stock 4 (BSS + Hep 8 ml) per bottle of OCM
1 aliquot of oocyte
maturation medium (
1
aliquot of stock 8 (gentamicin 1 ml) per aliquot of
1
aliquot of stock 3 (BSS 10 ml) per aliquot of
1
aliquot of stock 6 (Folltropin 125 ml) per aliquot of
1
aliquot of stock 10 (glutamine 1 ml) per aliquot of
200 ml of Stock 5 (estradiol) per aliquot of
1 ml
of Stock 2 (sodium pyruvate) per aliquot of
Laminar flow hood
1. The day before the collection of ovaries,
prepare and place at 4°C until use.
a) Saline + Pen/Strep -add 1 aliquot (10 ml) of
Stock 16: pen/strep to each liter of transport saline
b) OCM + Supplements - add supplements to
OCM as described
c)
2. Place saline (0.5 L/container) into two clean containers (Fig. 1). Place both containers in a thermos and leave at room temperature for use the next day.
Figure 1. This container is an example of one that is suitable for transportation of ovaries. Almost any leak-proof plastic container will work.
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