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A1: Konditionelle Immortalisierung von embryonalen Mäusezellen zur Untersuchung der Linienspezifiät

— abgelegt unter:

Projektleitung: Prof. Dr. F. Stewart / Dr. K. Anastassiadis

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Embryonic stem (ES) cells are characterised by their potential to give rise to a spectrum of differentiated cell types. The molecular mechanisms involved in maintenance of pluripotency or escape to lineage commitment are not understood. Our laboratories are working together on the proposition that pluripotency represents a naive, uncommitted, state of the epigenome and lineage commitment is, in part, a process of lineage specific epigenetic marking. Studies that explore epigenetic writing mechanisms in mouse ES cell differentiation and early development have been limited by our inability to obtain pure cell populations for biochemical analysis. To solve this problem, we developed a strategy for reversible, ligand regulated, cellular immortalization using SV40 Large T-antigen. Excellent Large T regulation in ES cells has been achieved and subsequently a mouse line has been established that also has excellent properties.

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Figure 1: Schematic presentation of the experimental approach. The expression of Large T-Antigen will be induced during differentiation of ES cells. Rare cell types will be isolated either by cellular cloning or by lineage selection and will be conditionally propagated. The same approach can be applied to adult stem cells isolated from transgenic mice.

The further exploration of this conditional immortalization strategy for the acquisition of rare and intermediate cell types is presented in this SFB for two reasons. First, to enhance our goal to study epigenetic mechanisms in lineage commitment. Second, to explore the possibility that the strategy can be applied to the conditional immortalization of precursor cell types that will enhance the research of SFB partners.