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Establishment of immortalized murine mesothelial cells and a novel mesothelioma cell line.

Blum W, Pecze L, Felley-Bosco E, Worthmüller-Rodriguez J, Wu L, Vrugt B, de Perrot M, Schwaller B - In Vitro Cell. Dev. Biol. Anim. (2015)

Bottom Line: All immortalized mesothelial clones consistently grow in DMEM supplemented with fetal bovine serum.Cells can be passaged for more than 40 times without any signs of morphological changes or a decrease in proliferation rate.RN5 cells are highly tumorigenic.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Fribourg, Route Albert-Gockel 1, 1700, Fribourg, Switzerland.

ABSTRACT
Mesothelial cells are susceptible to asbestos fiber-induced cytotoxicity and on longer time scales to transformation; the resulting mesothelioma is a highly aggressive neoplasm that is considered as incurable at the present time Zucali et al. (Cancer Treatment Reviews 37:543-558, 2011). Only few murine cell culture models of immortalized mesothelial cells and mesothelioma cell lines exist to date. We generated SV40-immortalized cell lines derived from wild-type (WT) and neurofibromatosis 2 (merlin) heterozygote (Nf2+/-) mice, both on a commonly used genetic background, C57Bl/6J. All immortalized mesothelial clones consistently grow in DMEM supplemented with fetal bovine serum. Cells can be passaged for more than 40 times without any signs of morphological changes or a decrease in proliferation rate. The tumor suppressor gene NF2 is one of the most frequently mutated genes in human mesothelioma, but its detailed function is still unknown. Thus, these genotypically distinct cell lines likely relevant for malignant mesothelioma formation are expected to serve as useful in vitro models, in particular to compare with in vivo studies in mice of the same genotype. Furthermore, we generated a novel murine mesothelioma cell line RN5 originating from an Nf2+/- mouse subjected to repeated crocidolite exposure. RN5 cells are highly tumorigenic.

No MeSH data available.


Related in: MedlinePlus

Morphology (a) and growth characteristics (b) of primary mesothelial cells isolated from WT and Nf2+/− mice, both grown in Connell’s medium. (a) Mesothelial cells from both genotypes show the typical “cobblestone-like” morphology. (b) Nf2+/− cells show slightly increased proliferation compared to WT cells; representative growth curves for n > 3 experiments are shown.
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Fig1: Morphology (a) and growth characteristics (b) of primary mesothelial cells isolated from WT and Nf2+/− mice, both grown in Connell’s medium. (a) Mesothelial cells from both genotypes show the typical “cobblestone-like” morphology. (b) Nf2+/− cells show slightly increased proliferation compared to WT cells; representative growth curves for n > 3 experiments are shown.

Mentions: Primary mesothelial cells isolated from WT and Nf2+/− mice were initially grown in Connell’s medium, a medium previously shown to be optimal for mesothelial cell growth (Connell and Rheinwald 1983). Growth of cells from both genotypes was faster then in the medium used for the further experiments, i.e., in DMEM supplemented with 10% FBS (data not shown). Primary mesothelial cells from WT and Nf2+/− mice grown in DMEM showed the typical cobblestone-like morphology similar as, e.g., the human mesothelioma cell line ZL55 consisting of cells with an epithelioid morphology used in previous studies (Blum and Schwaller 2013) (Fig. 1a). As shown before for WT primary mouse mesothelial cells (Robinson et al.2006), mesothelial cells from the two genotypes grew in vitro for approximately 8–10 passages, followed by a prolonged period of quiescence lasting for up to several weeks before finally dying.Figure 1.


Establishment of immortalized murine mesothelial cells and a novel mesothelioma cell line.

Blum W, Pecze L, Felley-Bosco E, Worthmüller-Rodriguez J, Wu L, Vrugt B, de Perrot M, Schwaller B - In Vitro Cell. Dev. Biol. Anim. (2015)

Morphology (a) and growth characteristics (b) of primary mesothelial cells isolated from WT and Nf2+/− mice, both grown in Connell’s medium. (a) Mesothelial cells from both genotypes show the typical “cobblestone-like” morphology. (b) Nf2+/− cells show slightly increased proliferation compared to WT cells; representative growth curves for n > 3 experiments are shown.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4539351&req=5

Fig1: Morphology (a) and growth characteristics (b) of primary mesothelial cells isolated from WT and Nf2+/− mice, both grown in Connell’s medium. (a) Mesothelial cells from both genotypes show the typical “cobblestone-like” morphology. (b) Nf2+/− cells show slightly increased proliferation compared to WT cells; representative growth curves for n > 3 experiments are shown.
Mentions: Primary mesothelial cells isolated from WT and Nf2+/− mice were initially grown in Connell’s medium, a medium previously shown to be optimal for mesothelial cell growth (Connell and Rheinwald 1983). Growth of cells from both genotypes was faster then in the medium used for the further experiments, i.e., in DMEM supplemented with 10% FBS (data not shown). Primary mesothelial cells from WT and Nf2+/− mice grown in DMEM showed the typical cobblestone-like morphology similar as, e.g., the human mesothelioma cell line ZL55 consisting of cells with an epithelioid morphology used in previous studies (Blum and Schwaller 2013) (Fig. 1a). As shown before for WT primary mouse mesothelial cells (Robinson et al.2006), mesothelial cells from the two genotypes grew in vitro for approximately 8–10 passages, followed by a prolonged period of quiescence lasting for up to several weeks before finally dying.Figure 1.

Bottom Line: All immortalized mesothelial clones consistently grow in DMEM supplemented with fetal bovine serum.Cells can be passaged for more than 40 times without any signs of morphological changes or a decrease in proliferation rate.RN5 cells are highly tumorigenic.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Fribourg, Route Albert-Gockel 1, 1700, Fribourg, Switzerland.

ABSTRACT
Mesothelial cells are susceptible to asbestos fiber-induced cytotoxicity and on longer time scales to transformation; the resulting mesothelioma is a highly aggressive neoplasm that is considered as incurable at the present time Zucali et al. (Cancer Treatment Reviews 37:543-558, 2011). Only few murine cell culture models of immortalized mesothelial cells and mesothelioma cell lines exist to date. We generated SV40-immortalized cell lines derived from wild-type (WT) and neurofibromatosis 2 (merlin) heterozygote (Nf2+/-) mice, both on a commonly used genetic background, C57Bl/6J. All immortalized mesothelial clones consistently grow in DMEM supplemented with fetal bovine serum. Cells can be passaged for more than 40 times without any signs of morphological changes or a decrease in proliferation rate. The tumor suppressor gene NF2 is one of the most frequently mutated genes in human mesothelioma, but its detailed function is still unknown. Thus, these genotypically distinct cell lines likely relevant for malignant mesothelioma formation are expected to serve as useful in vitro models, in particular to compare with in vivo studies in mice of the same genotype. Furthermore, we generated a novel murine mesothelioma cell line RN5 originating from an Nf2+/- mouse subjected to repeated crocidolite exposure. RN5 cells are highly tumorigenic.

No MeSH data available.


Related in: MedlinePlus