Materials

1. Recombinant human follicle-stimulating hormone, luteinizing hormone (LH), and chorionic gonadotrophin (CG; Ares Advanced Technologies, Norwell, MA; or recombinant monkey gonadotropins when available).

2. Antide (GnRH antagonist, Ares Advanced Technologies).

4. TH3 medium: HEPES-buffered TALP medium, containing 0.3% bovine serum albumin (BSA [6]). Prepare medium by adding the indicated amounts of each reagent (Sigma, St. Louis, MO) to 1 L of Milli-Q water.

NaCl

6.660 g

KCl

0.239 g

CaCl22H2O

0.294 g

MgCl2-6H2O

0.102 g

Na2HPO4

0.048 g

Glucose

0.900 g

Na lactate

1.87 mL

Phenol red

0.010 g

NaHCO3

0.168 g

Gentamicin sulfate

0.050 g

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