Cell Lines Derived From Feline Fibrosarcoma Display Unstable Chromosomal Aneuploidy and Additionally Centrosome Number Aberrations

Abstract

The purpose of the study was to evaluate clonality and presence of numerical chromosomal and centrosomal aberrations in 5 established feline fibrosarcoma cell lines and in a fetal dermal fibroblast cell line as a control. The clonality of all cell lines was examined using limited-dilution cloning. The number of chromosomes was counted in metaphase spreads. The immunocytochemical analysis of centrosome numbers was performed by indirect immunofluorescence using a monoclonal antibody that targets g-tubulin, a well-characterized component of centrosomes. Monoclonal cell populations could be established from all cell lines. In all feline fibrosarcoma cell lines, the number of chromosomes deviated abnormally from the normal feline chromosome number of 2n ¼ 38, ranging from 19 to 155 chromosomes per cell. Centrosome hyperamplification was observed in all 5 feline fibrosarcoma cell lines with a proportion of cells (5.7 to 15.2%) having more than 2 centrosomes. In the control cell line, only 0.6% of the cells had more than 2 centrosomes. In conclusion, the examinations revealed that centrosome hyperamplification occurs in feline fibrosarcoma cell lines. The feline fibrosarcoma cell lines possessed 10 to 25 times as many cells with centrosome hyperamplification as the control cell line. These observations suggest an association of numerical centrosome aberrations with karyotype instability by increasing the frequency of chromosome missegregation. The results of this study may be helpful for further characterization of feline fibrosarcomas and may contribute to the knowledge of cytogenetic factors that may be important for the pathogenesis of feline fibrosarcomas.

Enhancing Anti-Tumor Efficacy of Doxorubicin by Non-Covalent Conjugation to Gold Nanoparticles – In Vitro Studies on Feline Fibrosarcoma Cell Lines

“…FFS1WAW were derived in the Warsaw University of Life Sciences according to the protocol described in our previous studies. [ 39 ] FFS1, FFS3 and FFS5 cell lines were derived by Erlichsen et al[ 40 ] Cells were cultured in standard conditions (in an atmosphere of 5% CO 2 , 95% humidified air at 37°C) in DMEM-high glucose medium (Gibco BRL, Scotland) enriched with 10% heat-inactivated fetal bovine serum (FBS, Sigma Aldrich, USA) and antibiotics: Penicillin with Streptomycin (50 I.U./mL, Gibco BRL, Scotland), Amphotericin B (2.5 mg/mL, Gibco BRL, Scotland) and grown in tissue culture flasks (Becton Dickinson, USA). The medium was changed every 48–72 hours (depending on confluence of cells) in order to assure optimal conditions for cell growth.…”

Doxorubicin Conjugated to Glutathione Stabilized Gold Nanoparticles (Au-GSH-Dox) as an Effective Therapeutic Agent for Feline Injection-Site Sarcomas—Chick Embryo Chorioallantoic Membrane Study

“…The FFS1 and FFS3 cell lines were obtained from Justus Liebig Universitat (Giessen, Germany) [ 35 ]. Cells were cultured in a 5% CO 2 incubator with high glucose (4500 g/L) Dulbecco’s Modified Eagle Meddium (Gibco BRL, Life Technologies, Paisley, UK) enriched with heat-inactivated fetal bovine serum (FBS), penicillin-streptomycin (50 mL IU-1) and amphotericin B (2.5 mg·mL −1 ).…”

3D chick embryo chorioallantoic membrane model as an in vivo model to study morphological and histopathological features of feline fibrosarcomas

“…Feline fibrosarcoma cell lines (FFS1, FFS3) derived in Justus Liebig Universitat in Giessen (Germany) [41] were cultivated under standard aseptic conditions (5% CO 2 , 95% humidity and 37 °C) in Dulbeco’s Modified Eagle Medium (DMEM) with glucose (4500 mg/L) (Gibco BRL), enriched with 10% v /v heat inactivated fetal bovine serum (FBS), penicillin-streptomycin (50 mL IU-1), amphotericin B (2.5 mg/mL-1) (reagents obtained from Sigma Aldrich, USA). The medium was changed every 48–72 h, when cell confluence reached 75–80%.…”