Defining the genetic characteristics of human hepatic stellate cell line LX-2 by spectral karyotyping and short tandem repeat genotyping

R Weiskirchen; S Meurer; L Xu; S Friedman; C Bergmann

doi:10.1055/s-0032-1331955

Zeitschrift für Gastroenterologie, Table of Contents

Defining the genetic characteristics of human hepatic stellate cell line LX-2 by spectral karyotyping and short tandem repeat genotyping

R Weiskirchen ¹, S Meurer ¹, L Xu ², S Friedman ³, C Bergmann ⁴

¹RWTH Aachen University, Institute of Clinical Chemistry and Pathobiochemistry, Aachen, Germany
²Shanghai University of Traditional Chinese Medicine Shuguang Hospital, Institute of Liver Diseases, Shanghai, PR China
³Mount Sinai School of Medicine, Division of Liver Diseases, New York, USA
⁴Bioscientia, Center for Human Genetics, Ingelheim, Germany

Abstract

Objective: The hepatic cell line LX-2 was originally described in 2005 as a novel tool to study mechanisms of hepatic fibrogenesis and the testing of antifibrotic compounds [1]. It was established by immortalisation with the Simian Vacuolating Virus 40 (SV40) transforming (T) antigen and subsequent propagation in low serum conditions. Although this immortalized line is used in an increasing number of studies [2], a detailed genotypic characterisation that is mandatory for immortalized cell lines is still lacking. Design: In this study we attempt to describe the genetic characteristics and expression attributes of LX-2 cells to establish a reliable platform for explaining its cellular behaviour. Results: We established a single-locus short tandem repeat (STR) signature for the cell line and characterized the LX-2 karyotype by several cytogenetic and molecular cytogenetic techniques. Spectral karyotyping (SKY) revealed that LX-2 have marked heterogeneity with some chromosomal aberrations. However, despite some complexity and genetic variability, a few recurrent translocations have been identified that are specific for LX-2 and therefore suitable for proper identity testing. In addition, we found that this LX-2 cells express collagen type IV, collagen type I, fibronectin, vimentin, desmin, α-smooth muscle actin, connective tissue growth factor (CTGF), Id2, p21 and the complete set of TGF-β receptors including TβRI, TβRII and the accessory receptor endoglin (CD105) supporting the notion that this cell line originated from hepatic stellate cells. Conclusions: Our study provides criteria for cellular authentication of LX-2, and offers functional clues underlying its phenotypic and genotypic features. The provided information will increase the attraction for LX-2 cells in liver research and allow easy cellular authentication to avoid potential false experimental outcomes and scientific misinterpretation in studies in which LX-2 are employed.

References:

[1] Xu L et al. GUT 2005;54:142–151.

[2] Herrmann J et al. J Cell Mol Med 2007;11:704–722.