Optimization of Three-Dimensional Culture Conditions of HepG2 Cells with Response Surface Methodology Based on the VitroGel System.

Jing Bo WANG; Wen QIN; Zhuo YANG; Shi SHEN; Yan MA; Li Yuan WANG; Qin ZHUO; Zhao Long GONG; Jun Sheng HUO; Chen CHEN

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Optimization of Three-Dimensional Culture Conditions of HepG2 Cells with Response Surface Methodology Based on the VitroGel System.

Author: Jing Bo WANG ¹ ; Wen QIN ¹ ; Zhuo YANG ¹ ; Shi SHEN ¹ ; Yan MA ¹ ; Li Yuan WANG ¹ ; Qin ZHUO ¹ ; Zhao Long GONG ¹ ; Jun Sheng HUO ¹ ; Chen CHEN ¹
Author Information

1. National Institute for Nutrition and Health, Chinese Centre for Disease Control and Prevention, Beijing 100050, China.
Publication Type:Journal Article
Keywords: Hydrogel; Response surface methodology; The three-dimensional culture
MeSH: Albumins; Cell Culture Techniques/methods*; Hep G2 Cells; Humans; Hydrogels
From: Biomedical and Environmental Sciences 2022;35(8):688-698
CountryChina
Language:English
Abstract: OBJECTIVE:This study optimizes three-dimensional (3D) culture conditions of HepG2 using response surface methodology (RSM) based on the VitroGel system to facilitate the cell model in vitro for liver tissues.
METHOD:HepG2 cell was 3D cultured on the VitroGel system. Cell viability was detected using Cell Counting Kit-8 (CCK-8) assay of HepG2 lived cell numbers. The proliferation of HepG2 cell and clustering performance was measured via fluorescence staining test. Albumin concentration in the culture medium supernatant as an index of HepG2 cell biological function was measured with ELISA kit. Independent factor tests were conducted with three key factors: inoculated cell concentration, cultured time, and dilution degree of the hydrogel. The preliminary results of independent factor tests were used to determine the levels of factors for RSM.
RESULT:The selected optimal culture conditions are as follows: concentration of inoculated cells was 4.44 × 10 ⁵/mL, culture time was 4.86 days, and hydrogel dilution degree was 1:2.23. The result shows that under optimal conditions, the predicted optical density (OD) value of cell viability was 3.10 and measured 2.978 with a relative error of 3.94%.
CONCLUSION:This study serves as a reference for the 3D HepG2 culture and constructs liver tissues in vitro. Additionally, it provides the foundation for repeated dose high-throughput toxicity studies and other scientific research work.