In vitro screening test for mutagenicity and genotoxicity during spermatogenesis

A serum-free media formulation for culturing Xenopus testis explants comprising one member selected from the group consisting of medium 1 and medium 2:

• ______________________________________ Serum-Free Media Formulations (mg/L) ______________________________________ Medium 1:
• NaCl 3786 Proline 60 KCl 186 Serine 240 Na.sub.2 HPO.sub.4.12H.sub.2 O 358 Threonine 70 CaCl.sub.2.2H.sub.2 O 147 Tryptophan 20 MgCl.sub.2.6H.sub.2 O 102 Tyrosine 40 MgSO.sub.4.7H.sub.2 O 123 Valine 60 FeSO.sub.4 0.834 Glucose 1000 HEPES (pH 7.4) 3576 Oxaloacetate 152 Alanine 130 Sodium Pyruvate 110 Arginine-HCl 140 Bovine Serum Albumin 1000 Aspartate-Mg 450 (fatty acid-free) Cysteine-HCl.H.sub.2 O 20 Transferrin (human) 10 Cystine 10 Basal Medium Eagles (1X) Vitamin Mix Glutamate-HCl 1200 Ascorbate 50 Glutamine 292 DL-a-Tocopherol 0.2 Glycine 60 Retinol 0.029 Histidine-HCl.H.sub.2 O 60 Bovine Insulin (Zn) 10 Isoleucine 50 Dihydrotestosterone 0.03 Leucine 80 Testosterone 0.03 Lysine-HCl 280 Follicle-Stimulating 5 Hormone Methionine 50 Antibiotic- (1X) Phenylalanine 50 Antimycotic Mix (100 units/ml penicillin;
• 100 mcg/ml streptomycin sulfate;
• 0.25 mcg/ml Fungizone) Medium 2:
• Medium 1 - not containing tocopherol, ascorbate and testosterone, supplemented with linoleic acid (5 .mu.g/ml). ______________________________________

A method for culturing Xenopus testis explants in vitro such that cells at all stages of a spermatogenic cycle are produced comprising culturing Xenopus testis fragments in one serum-free medium selected from the group consisting of medium 1 and medium 2:

• ______________________________________ Serum-Free Media Formulations (mg/L) ______________________________________ Medium 1:
• NaCl 3786 Proline 60 KCl 186 Serine 240 Na.sub.2 HPO.sub.4.12H.sub.2 O 358 Threonine 70 CaCl.sub.2.2H.sub.2 O 147 Tryptophan 20 MgCl.sub.2.6H.sub.2 O 102 Tyrosine 40 MgSO.sub.4.7H.sub.2 O 123 Valine 60 FeSO.sub.4 0.834 Glucose 1000 HEPES (pH 7.4) 3576 Oxaloacetate 152 Alanine 130 Sodium Pyruvate 110 Arginine-HCl 140 Bovine Serum Albumin 1000 Aspartate-Mg 450 (fatty acid-free) Cysteine-HCl.H.sub.2 O 20 Transferrin (human) 10 Cystine 10 Basal Medium Eagles (1X) Vitamin Mix Glutamate-HCl 1200 Ascorbate 50 Glutamine 292 DL-a-Tocopherol 0.2 Glycine 60 Retinol 0.029 Histidine-HCl.H.sub.2 O 60 Bovine Insulin (Zn) 10 Isoleucine 50 Dihydrotestosterone 0.03 Leucine 80 Testosterone 0.03 Lysine-HCl 280 Follicle-Stimulating 5 Hormone Methionine 50 Antibiotic- (1X) Phenylalanine 50 Antimycotic Mix (100 units/ml penicillin;
• 100 mcg/ml streptomycin sulfate;
• 0.25 mcg/ml Fungizone) Medium 2:
• Medium 1 - not containing tocopherol, ascorbate and testosterone, supplemented with linoleic acid (5 .mu.g/ml). ______________________________________
• under an atmosphere of air and at a temperature in a range of from about 20.degree. to about 24.degree. C.