Determining population doubling time and the appropriate number of HepG2 cells for culturing in 6-well plate
Journal Title: International Research Journal of Applied and Basic Sciences - Year 2016, Vol 10, Issue 3
Abstract
Background: HepG2 is a widely used cell line in experiments of 30 years. These cells have been obtained from a 15 year old white man with well-differentiated hepatocellular carcinoma (HCC). HepG2 cells have epithelial morphology and secrets a lot of plasma proteins. Objectives: The aim of this study was determining the viability and the growth rate of HepG2 cells at 24, 48 and 72 h and determining the appropriate cell number to seed into 6-well plate. Materials and methods: HepG2 cells were cultured in complete medium. 1×106, 1.2×106, 1.5×106 and 1.7×106 cells were seeded in each well of 6-well plate. Cell confluency percentage and population doubling time (PDT) and viability of them were calculated. Results: The counfluency of cells were 30-40 % at 24 h, 60-70 % at 48 h and 70-80 % at 72 h. population doubling time was 48 h in group with 1.2×106 cells. Conclusions: Our results showed that the most suitable number of HepG2 cells for 6-well plate is 1.2×106, because of accessibility to medium and each other. The cells doubled after 48 h and increased 2.5 times after 72 h that was consistent with other studies.
Authors and Affiliations
Marjan Norouzzadeh| Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetic, Tehran University of Medical Sciences, Tehran, Iran., Yas Kalikias| Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetic, Tehran University of Medical Sciences, Tehran, Iran., Zinat Mohamadpur| Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetic, Tehran University of Medical Sciences, Tehran, Iran., Loghman Sharifi| Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetic, Tehran University of Medical Sciences, Tehran, Iran., Maryam Mahmoudi*| Department of Cellular and Molecular Nutrition, School of Nutritional Sciences and Dietetic, Tehran University of Medical Sciences, Tehran, Iran. email:mahmoodi_maryam@yahoo.com
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