<p>There is an increasing demand for primary human hepatocytes for toxico-pharmacological studies and for exploring their role in liver cell transplantation. During the whole process of isolation, culture and cryopreservation hepatocytes are subjected to various stress. All these factors will lead to the impairment in the metabolic capability of the hepatocytes post isolation. Hepatocyte isolation is an expensive and resource driven procedure. Thus, it is important to consider all the factors before we subject the specimen for isolation of cells. By considering the factors that influence hepatocyte isolation, hepatocytes of good quality can be obtained. With recommendations on the ideal parameters for isolation being made the researcher shall have a knowledge on what critical factors need to be taken care of during hepatocyte isolation. There will be an opportunity to modify critical factors if possible, during the procurement of specimen by working in tandem with the clinicians. The specimen which shall be deemed un fit when two or more factors have exceeded the critical limits can be rejected.</p>