Production of TNF-α and IL-10 by HepG2 and THP-1 cells differs depending on whether ethanol conditioned media is derived from 3D vs. 2D cultured HepG2 cells.

Abstract

ID 16411 Poster Board 244 Background: One of the difficulties in cancer research is bridging the gap between human and animal tumor models. One proposed method of improving the screening of cancer-promoting toxins and novel cancer therapies is the development of pre-clinical 3D human culture models. We are currently developing 3D human hepatocellular carcinoma (HCC) culture models using the standardized HepG2 liver cancer cell line as a template. Ethanol is being used as a test agent to generate conditioned media (CM) from 2D adherent vs. 3D HepG2 suspension spheroid cultures. The conditioned media is then applied to human monocytes to determine differences in TNF-a (proinflammatory) and IL-10 (anti-inflammatory) cytokine induction.Objectives: The experimental goals were to determine differences in the production of TNF-a and IL-10 between 2D and 3D cultured HepG2 cells, and by THP-1 monocytes treated with conditioned media (CM), +/-ethanol, derived from 2D and 3D HepG2 cells.Methods: HepG2 cells were grown in both 2D (adherent) and 3D (suspension) cultures to produce two different patterns of cell growth (Fig. 1). The cells were then switched to fetal bovine exosome-free media and grown with and without 100 mM ethanol to produce CM. Human monocytes (THP-1 cells) were then cultured in the presence of CM. Cell viability and production of TNF-a and IL-10 were measured using PrestoBlue™ and Lumit™ immunoassays.