ISSN 2456-0235

International Journal of Modern Science and Technology


​​​​​​​August 2019, Vol. 4, No. 8, pp 212-216. 

​​Optimization of Protocol for Maintaining and Freezing Caco-2 Cells

Rasika Pawar¹, Siddhivinayak Barve², Minal Joshi², Pranjali Dhawal², Bela Nabar¹*
¹Department of Microbiology, Smt. Chandibai Himathmal Mansukhani College, Ulhasnagar, India.
²Scientific Research Center, Vinayak Ganesh Vaze College, Mulund, Mumbai, India.

​​*Corresponding author’s e-mail:


Human Caco-2 cells are used as model of intestine for absorption studies. However standardization of growth conditions is challenging for these cells as it requires a long growth period of around twenty one days for most absorption associated studies. In this study we have standardized various growth associated parameters and procedures related to Caco-2 cell cultures. The Caco-2 protocols are recently developed in our laboratory include, the time required for trypsinization of cell  in correlation with the age of subculture, freezing mixture type, quantity of DMSO and seeding density of cells while freezing. The cells are also seeded at low density as compared to present in literature which gave better results for a 21 day old differentiated flask. Since the Caco-2 cell line is integral part of absorption and in vitro toxicology tests, standardized protocol inclusive of all the detailed procedures for growing and differentiating Caco-2 cells would help researchers in maintaining and preserving the cells.

Keywords: Caco-2; Trypsinization; Seed density; Freezing; Thawing.


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