Leishmaniasis, a parasitic disease caused by the protozoan Leishmania is endemic in many parts of tropic and sub tropic areas of the world. In vitro cultivation of parasite plays an important role in the study and treatment of disease. Traditionally media available do not meet the requirement for the bulk cultivation of Leishmania parasites, it requires fetal calf serum (FCS), that is very expensive and not easily available in the market. It is reported that human urine has stimulatory effect on the growth of Leishmania while cultured in vitro. We undertook a detailed study of such an effect in old world Leishmania isolates causing cutaneous leishmaniasis. Different concentrations of urine are tried and it is found that 1% sterile urine in our undefined medium 1999 supplemented with foetal calf serum gives the maximum growth. It is observed that dated urine has less stimulatory effect on the growth of the parasites as compared to fresh urine. The effect of the urine of different groups is also observed and it is found that the urine of 60 year gives maximum growth.