Research
Comparative Proteomic Phenotyping of Cell Lines and Primary Cells to Assess Preservation of Cell Type-specific Functions

https://doi.org/10.1074/mcp.M800258-MCP200Get rights and content
Under a Creative Commons license
open access

Biological experiments are most often performed with immortalized cell lines because they are readily available and can be expanded without limitation. However, cell lines may differ from the in vivo situation in important aspects. Here we introduce a straightforward methodology to compare cell lines to their cognate primary cells and to derive a comparative functional phenotype. We used SILAC (stable isotope labeling by amino acids in cell culture) for quantitative, mass spectrometry-based comparison of the hepatoma cell line Hepa1–6 with primary hepatocytes. The resulting quantitative proteome of 4,063 proteins had an asymmetric distribution, with many proteins down-regulated in the cell line. Bioinformatic analysis of the quantitative proteomics phenotypes revealed that Hepa1–6 cells were deficient in mitochondria, reflecting re-arrangement of metabolic pathways, drastically up-regulate cell cycle-associated functions and largely shut down drug metabolizing enzymes characteristic for the liver. This quantitative knowledge of changes provides an important basis to adapt cell lines to more closely resemble physiological conditions.

Cited by (0)

Published, MCP Papers in Press, October 23, 2008, DOI 10.1074/mcp.M800258-MCP200

This work was supported by HepatoSys, by Interaction Proteome, a 6th framework program of the European Union, and by the Max-Planck Society.

The on-line version of this article (available at http://www.mcponline.org) contains supplemental material.

§

Both authors contributed equally to this work.