Abstract: Cichorium intybus a member of family Asteraceae, has been traditionally used as liver protectant and claimed as renal protective and antiinflammatory. It contains many essential lipids, vitamins and a variety of sugar. For routine primary culture and for optimum growth in several secondary cultures of animal cell serum is frequently used as a source of nutrients. Isolation of serum from fetal bovine being barbarous process as well as its use in culture media is also associated with several drawbacks. Available animal free media are having their own shortcomings. The objective of this study is to evaluate the plant’s methanolic extract for in vitro cell growth supporting activity on rat hepatocyte primary culture. Plant extract was evaluated at five different concentrations in 96 well plates. At highest concentration of plant extract (50 μg mL-1) cell viability was found to be 66.83%, whereas, at lowest concentration (10 μg mL-1) cell viability was 24.12%. Plant extract was also found to overcome toxic effect of DMSO. Hence this work can be used for further characterization of the particular compound/s support the growth of rat primary hepatocyte culture as well as if cells of other origin and types. It will help in the formulation of novel animal cell culture medium based on plant origin/components, at least for the rat hepatocyte primary culture.