Background: Results of artificial insemination (AI) still does not exceed 20-25% because of different factors one of them is male factor infertility. Therefore, the global companies concern on the culture media that used for activation of semen samples of infertile men. However, till now, there are many semen samples did not response to these media leading to failure in AI. The objective of this study is to improve certain sperm function parameters in vitro of asthenoteratospermic patients by using novel combination motility stimulants substances that improved medium to be used for assisted reproductive technologies (such as artificial insemination) in future. One hundred asthenozoospermic patients are involved in this study. They are divided randomly into equal four groups. Each semen sample had been divided into two parts. One part was considered as a control by using Ham’s F10 medium and the other part is considered as treated group by adding the following substances in combination; Pentoxifylline (PF) 1mg/ml, Progesterone (P) 0.409 mg/ml and Glycyrrhiza glabra (Gg) 1mg/ml in the following groups. Group 1:PF and Gg, Group2: PF and P,Group3: Gg and P, and Group4:PF,Gg and P. Sperm function parameters are examined before and following in vitro preparation using wash and spin technique. The results revealed highly significant increment (P<0.01) in the percentage of totalactive sperm motility grades (a+b) andforward progressive sperm motility grade (a) with a significant improvement (P<0.05) in the percentage of progressive sperm motility grade (b) and morphologically normal sperm when using PFand Gg medium in comparison with control medium and other combination medium following 10, 30 and 60 minutes incubation The pentoxifylline is a phosphodiestrase inhibitor of the methylxanthine group which inhibit the degradationof cAMP of the sperm and increase the influx of Ca++ inside the sperm cell leading to increase the sperm motility. The Gg is a herbal plant th