With the numerous improvements in cereal tissue and wheat anther culture, it is necessary to determine which of the improvements should be combined for optimal response. This study was conducted using one highly responsive cultivar of wheat (Triticum aestivum L. cv. Pavon 76) to test the effectiveness of pre-culture cold treatment (0 or 3-7 days at 5oC) of anthers, five initiation basal media, and various changes in Murashige-Skoog regeneration media. A cold pre-culture treatment was inhibitory for all initiation media for embryoid initiation. Of the initiation media, P1, 85D12, and N6 were similar for embryoid initiation (0.80 to 0.90 embryoids/anther) without a cold pre-culture treatment. Plant regeneration was improved by the addition of amino acids or glucose, increased sucrose concentration, filter sterilizing the medium, and altering plant growth regulator concentrations. P1 medium which is normally used for embryoid initiation was also beneficial for improving plant regeneration. Ethylene inhibitors were generally not beneficial.