Department of Virology, Post Graduate Institute of Medical Education & Research, Chandigarh, India.
Abstracts of the papers presented at the XVIII National Conference of Indian Virological Society at Post Graduate Institute of Medical Education and Research, Chandigarh, India, December 11–13, 2008.
Virus isolation is an essential component of both diagnostic and research virus laboratory. Presently cell culture is the most popular method of virus propagation. The maintenance of cell lines at low passage level is important in order to preserve its sensitivity. The need of high cost liquid nitrogen for storage of cell lines is however one of the limiting factor for its widespread use in developing countries. The present study was therefore carried out to standardize the preservation of Vero and Hep 2 cell lines in deep freezer (−850 c) for 6 months. The experiment was carried out in Vero and Hep2 cell lines. Both the cell lines were grown in multiple cell culture flasks. After formation of monolayer, cells were collected and fixed number of cells were preserved in 12 vials in each batch at −85°c deep freezer. Two vials were revived from each batch at one month intervals. The percentage of cell adherence, pattern of cell growth, morphology of cells and time taken for monolayer formation were observed and recorded. The process was continued till 6 months of preservation and were repeated three times with each of Vero and Hep2 cell lines. Both the cell lines could be revived with healthy cells and monolayer was formed within 7-10 days, after storage at −85°c for 4months. The present study highlights the utility of -85°c deep freezer as a cheaper alternative to liquid N2 for preservation of these cell lines at least up to 4 months.
