TY  - JOUR
A1  - Nanda, Indrajit
A1  - Schröder, Sarah K.
A1  - Steinlein, Claus
A1  - Haaf, Thomas
A1  - Buhl, Eva M.
A1  - Grimm, Domink G.
A1  - Weiskirchen, Ralf
T1  - Rat hepatic stellate cell line CFSC-2G: genetic markers and short tandem repeat profile useful for cell line authentication
JF  - Cells
N2  - Hepatic stellate cells (HSCs) are also known as lipocytes, fat-storing cells, perisinusoidal cells, or Ito cells. These liver-specific mesenchymal cells represent about 5% to 8% of all liver cells, playing a key role in maintaining the microenvironment of the hepatic sinusoid. Upon chronic liver injury or in primary culture, these cells become activated and transdifferentiate into a contractile phenotype, i.e., the myofibroblast, capable of producing and secreting large quantities of extracellular matrix compounds. Based on their central role in the initiation and progression of chronic liver diseases, cultured HSCs are valuable in vitro tools to study molecular and cellular aspects of liver diseases. However, the isolation of these cells requires special equipment, trained personnel, and in some cases needs approval from respective authorities. To overcome these limitations, several immortalized HSC lines were established. One of these cell lines is CFSC, which was originally established from cirrhotic rat livers induced by carbon tetrachloride. First introduced in 1991, this cell line and derivatives thereof (i.e., CFSC-2G, CFSC-3H, CFSC-5H, and CFSC-8B) are now used in many laboratories as an established in vitro HSC model. We here describe molecular features that are suitable for cell authentication. Importantly, chromosome banding and multicolor spectral karyotyping (SKY) analysis demonstrate that the CFSC-2G genome has accumulated extensive chromosome rearrangements and most chromosomes exist in multiple copies producing a pseudo-triploid karyotype. Furthermore, our study documents a defined short tandem repeat (STR) profile including 31 species-specific markers, and a list of genes expressed in CFSC-2G established by bulk mRNA next-generation sequencing (NGS).
KW  - liver
KW  - extracellular matrix
KW  - hepatic stellate cell
KW  - myofibroblast
KW  - fibrosis
KW  - stress fibers
KW  - spectral karyotyping
KW  - rhodamine–phalloidin stain
KW  - next-generation sequencing
KW  - STR profile
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-288067
SN  - 2073-4409
VL  - 11
IS  - 18
ER  - 
TY  - JOUR
A1  - Nanda, Indrajit
A1  - Steinlein, Claus
A1  - Haaf, Thomas
A1  - Buhl, Eva M.
A1  - Grimm, Domink G.
A1  - Friedman, Scott L.
A1  - Meurer, Steffen K.
A1  - Schröder, Sarah K.
A1  - Weiskirchen, Ralf
T1  - Genetic characterization of rat hepatic stellate cell line HSC-T6 for in vitro cell line authentication
JF  - Cells
N2  - Immortalized hepatic stellate cells (HSCs) established from mouse, rat, and humans are valuable in vitro models for the biomedical investigation of liver biology. These cell lines are homogenous, thereby providing consistent and reproducible results. They grow more robustly than primary HSCs and provide an unlimited supply of proteins or nucleic acids for biochemical studies. Moreover, they can overcome ethical concerns associated with the use of animal and human tissue and allow for fostering of the 3R principle of replacement, reduction, and refinement proposed in 1959 by William M. S. Russell and Rex L. Burch. Nevertheless, working with continuous cell lines also has some disadvantages. In particular, there are ample examples in which genetic drift and cell misidentification has led to invalid data. Therefore, many journals and granting agencies now recommend proper cell line authentication. We herein describe the genetic characterization of the rat HSC line HSC-T6, which was introduced as a new in vitro model for the study of retinoid metabolism. The consensus chromosome markers, outlined primarily through multicolor spectral karyotyping (SKY), demonstrate that apart from the large derivative chromosome 1 (RNO1), at least two additional chromosomes (RNO4 and RNO7) are found to be in three copies in all metaphases. Additionally, we have defined a short tandem repeat (STR) profile for HSC-T6, including 31 species-specific markers. The typical features of these cells have been further determined by electron microscopy, Western blotting, and Rhodamine-Phalloidin staining. Finally, we have analyzed the transcriptome of HSC-T6 cells by mRNA sequencing (mRNA-Seq) using next generation sequencing (NGS).
KW  - liver
KW  - extracellular matrix
KW  - hepatic stellate cell
KW  - myofibroblast
KW  - fibrosis
KW  - in vitro model
KW  - SKY analysis
KW  - phalloidin stain
KW  - next generation sequencing
KW  - STR profile
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-275178
SN  - 2073-4409
VL  - 11
IS  - 11
ER  -