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Characterization of the murine myeloid precursor cell line MuMac-E8.

ABSTRACT: Starting point for the present work was the assumption that the cell line MuMac-E8 represents a murine cell population with stem cell properties. Preliminary studies already pointed to the expression of stem-cell associated markers and a self-regenerative potential of the cells. The cell line MuMac-E8 should be examined for their differential stage within stem cell hierarchy. MuMac-E8 cells were derived from a chimeric mouse model of arthritis. It could be shown that MuMac-E8 cells express mRNA of some genes associated with pluripotent stem cells (Nanog, Nucleostemin), of genes for hematopoietic markers (EPCR, Sca-1, CD11b, CD45), for the mesenchymal marker CD105 and of genes for the neural markers Pax-6 and Ezrin. In methylcellulose and May-Grünwald-Giemsa staining, hematopoietic colonies were obtained but the hematopoietic system of lethally irradiated mice could not be rescued. Osteogenic differentiation was not detectable. Thus, it became evident that MuMac-E8 represents not a stem cell line. However, MuMac-E8 cells expressed several myeloid surface markers (i.e. CD11b, F4/80, CD14, CD64), showed phagocytosis and is capable of producing nitric oxide. Thus, this cell line seems to be arrested an advanced stage of myeloid differentiation. Adherence data measured by impedance-based real-time cell analysis together with cell morphology data suggested that MuMac-E8 represents a new macrophage precursor cell line exhibiting weak adherence. This cell line is suitable as an in-vitro model for testing of macrophage functions. Moreover, it might be also useful for differentiation or reprogramming studies.

SUBMITTER: Fricke S

PROVIDER: S-EPMC4278753 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Characterization of the murine myeloid precursor cell line MuMac-E8.

Fricke Stephan S Pfefferkorn Cathleen C Wolf Doris D Riemschneider Sina S Kohlschmidt Janine J Hilger Nadja N Fueldner Christiane C Knauer Jens J Sack Ulrich U Emmrich Frank F Lehmann Jörg J

PloS one 20141229 12

Starting point for the present work was the assumption that the cell line MuMac-E8 represents a murine cell population with stem cell properties. Preliminary studies already pointed to the expression of stem-cell associated markers and a self-regenerative potential of the cells. The cell line MuMac-E8 should be examined for their differential stage within stem cell hierarchy. MuMac-E8 cells were derived from a chimeric mouse model of arthritis. It could be shown that MuMac-E8 cells express mRNA ...[more]

PMID: 25546418

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Project description:Type 1 diabetes mellitus (T1DM) is caused by the selective destruction of insulin-producing ?-cells. This process is mediated by cells of the immune system through release of nitric oxide, free radicals and pro-inflammatory cytokines, which induce a complex network of intracellular signalling cascades, eventually affecting the expression of genes involved in ?-cell survival.The aim of our study was to investigate possible mechanisms of resistance to cytokine-induced ?-cell death. To this purpose, we created a cytokine-resistant ?-cell line (?-TC3R) by chronically treating the ?-TC3 murine insulinoma cell line with IL-1? + IFN-?. ?-TC3R cells exhibited higher proliferation rate and resistance to cytokine-mediated cell death in comparison to the parental line. Interestingly, they maintained expression of ?-cell specific markers, such as PDX1, NKX6.1, GLUT2 and insulin. The analysis of the secretory function showed that ?-TC3R cells have impaired glucose-induced c-peptide release, which however was only moderately reduced after incubation with KCl and tolbutamide. Gene expression analysis showed that ?-TC3R cells were characterized by downregulation of IL-1? and IFN-? receptors and upregulation of SOCS3, the classical negative regulator of cytokines signaling. Comparative proteomic analysis showed specific upregulation of 35 proteins, mainly involved in cell death, stress response and folding. Among them, SUMO4, a negative feedback regulator in NF-kB and JAK/STAT signaling pathways, resulted hyper-expressed. Silencing of SUMO4 was able to restore sensitivity to cytokine-induced cell death in ?-TC3R cells, suggesting it may play a key role in acquired cytokine resistance by blocking JAK/STAT and NF-kB lethal signaling.In conclusion, our study represents the first extensive proteomic characterization of a murine cytokine-resistant ?-cell line, which might represent a useful tool for studying the mechanisms involved in resistance to cytokine-mediated ?-cell death. This knowledge may be of potential benefit for patients with T1DM. In particular, SUMO4 could be used as a therapeutical target.

Dataset Information

Characterization of the murine myeloid precursor cell line MuMac-E8.

Publications

Characterization of the murine myeloid precursor cell line MuMac-E8.

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