Tissue and Cell Culture Reagents

1. Phosphate-buffered saline (PBS) without calcium/magnesium (Gibco).

2. Glasgow minimal essential medium (GMEM): Comprises IX GMEM supplemented with 10% fetal calf serum (FCS), 2 mM glutamine, 1 mM sodium pyruvate, 1X nonessential amino acids.

4. 200 mM Glutamine (Gibco).

5. 100 mM Sodium pyruvate (Gibco).

6. 100X MEM nonessential amino acids (Gibco).

7. FCS (Globe Farm, Guilford, or equivalent supplier).

8. Trypsin-EDTA: 1X solution (Gibco).

9. Gelatin (Sigma). Made to 0.1% solution in water and autoclaved.

10. Dimethyl sulfoxide (DMSO; Sigma).

11. Synthetic oviduct fluid (SOF; per 100 mL): 5.83 g NaCl, 0.534 g KCl, 0.162 g KH2PO4, 0.1 g MgCl2 2H2O, 0.616 g Na lactate, 0.27 g glucose, 0.06 g of penicillin, 0.05 g steptomycin, 2.1 g NaHCO3, 0.01 g Phenol Red, 0.36 g pyruvate,

2.52 g CaCl2H2O, 1.46 g gluthamine, 0.8 g bovine serum albumin, 1.0 mL nonessential amino acids, and 2.0 mL essential amino acids.

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