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.
Highlights
In recent years the investigation and characterization of new stem cell lines for improvement of cellular therapies came strongly into the focus of science
In addition to immunophenotyping of MuMac-E8 cells by flow cytometry, the principal objective of this work was the establishment of quantitative real-time polymerase chain reaction (PCR) assays for gene expression analysis of stem-cell- and lineage-associated markers using the Universal Probe Library (UPL) method
Cell status is represented by a dimensionless parameter termed Cell Index (CI) which is derived as the relative change in measured electrical impedance
Summary
In recent years the investigation and characterization of new stem cell lines for improvement of cellular therapies came strongly into the focus of science. Because of their great potential they are a beacon of hope in areas of transplantation and regenerative medicine. IL-4 is a potent suppressor of Th1-mediated mechanisms, which are still thought to play a role in various autoimmune diseases [3, 4] For this purpose, IL-4-transfected murine fibroblasts (NIH-3T3BMG-Neo-IL-4) [5] were injected into the affected knee joint of mice three days after intraarticular application of human RA fibroblasts. The knee joint swelling was observed over 6 weeks In this process the RA fibroblasts induced murine/human SCID arthritis worsened massively by injection of 3T3-IL-4 fibroblasts. In all three control groups, there was observed only a transient moderate swelling of the treated knee joint (Lehmann, J. et al unpublished data)
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