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Physical (seeding density, incubation period, incubation temperature and rolling speed) and chemical (amount of cell culture growth medium and fetal calf serum) factors were optimized for enhancing biomaö production of Baby Hamster Kidney cells (BHK-21) cells in roller bottle. The roller culture bottles (480 cm[sup 2]) were used for the propagation of the cells. Seeding of 10 million cells per bottle showed complete monolayer with maximum cell density (4.7 ×10[sup 7]) within 60 h that was significantly higher than those of other seeding densities of the cells (P< 0.05). The BHK-21 cells grew well at 35oC and 37oC within 60 h post incubation and formed confluent monolayer. However, the cell count declined significantly when the roller bottles were incubated at 39oC (P< 0.05). The bottles kept at rolling speed of 3 rpm yielded maximum count of the cells while higher or lower rotation speed showed adverse effects on the cell growth. Feeding of the cells with 100 mL of the growth medium (5% fetal calf serum) showed optimum growth of the cells. It is concluded that BHK-21 cells in roller bottle (480 cm[sup 2]) grew well and showed more than 4.7×10[sup 7] cell count/bottle under optimized conditions such as 10 million seeding cell density, incubation time 60 h, incubation temperature 37oC, 3 rpm, and 100 mL growth medium. © 2014 Friends Science Publishers [ABSTRACT FROM AUTHOR]