Continuous live quantification of molecular pluripotency control
Final Report Abstract
Goal of this project is the development of microscopy and software tools, and of ESC reporter lines to allow the live long-term quantification of cell behavior and dynamic transcription factor expression in pluripotent cells. These tools are to be used to quantify pluripotency transcription factor expression over time and with and without molecular manipulation to learn about the dynamics of transcription factor networks underlying pluripotency control. The project proceeds as planned, and already allowed important novel insights into the behavior of transcription factor networks, and corrected several prominent assumptions in the field. We found unexpected kinetics, memory, plasticity and lack of co-regulation of different core pluripotency transcription factors. Numerous tools for the timed and titratable manipulation of pluripotency transcription factors have been developed and are now used to further analyze the topology, dynamics and robustness of the pluripotency transcription factor network. The novel type of continuous single cell quantitative expression data is used for the development and falsification of refined computational models of pluripotency control.
Publications
- (2009). Continuous single-cell imaging of blood generation from haemogenic endothelium. Nature, 457: 896 – 900
Eilken HM, Nishikawa SI and Schroeder T
(See online at https://doi.org/10.1038/nature07760) - (2009). Hematopoietic cytokines can instruct lineage choice. Science, 325: 217 – 218
Rieger MA, Hoppe PS, Smejkal BM, Eitelhuber AC and Schroeder T
(See online at https://doi.org/10.1126/science.1171461) - (2010). Directing Astroglia from the Cerebral Cortex into Subtype Specific Functional Neurons. PLoS Biology, 8: e1000373
Heinrich C, Blum R, Gascon S, Masserdotti G, Tripathi P, Sánchez R, Tiedt S, Schroeder T, Götz M and Berninger B
(See online at https://doi.org/10.1371/journal.pbio.1000373) - (2011) Lentiviral vector design and imaging approaches to visualize the early stages of cellular reprogramming. Molecular Therapy, 19: 782-789
Warlich E, Kühle J, Cantz T, Brugman M, Maetzig T, Galla M, Filipczyk AA, Foerster R, Klump H, Schöler HR, Baum C, Schroeder T and Schambach A
(See online at https://doi.org/10.1038/mt.2010.314) - (2011). Continuous live imaging of adult neural stem cell division and lineage progression in vitro. Development, 138: 1057-1068
Costa M, Ortega F, Brill M, Beckervordersandforth-Bonk R, Petrone C, Schroeder T, Götz M and Berninger B
(See online at https://doi.org/10.1242/dev.061663) - (2011). Functionally Defined Substates within the Human Embryonic Stem Cell Compartment. Stem Cell Research, 7:145-153
Tonge PD, Shigeta M, Schroeder T, Andrews PW
(See online at https://doi.org/10.1016/j.scr.2011.04.006) - (2011). Generation of subtype-specific neurons from postnatal astroglia of the mouse cerebral cortex. Nature Protocols, 6: 214-228
Heinrich C, Gascón S, Masserdotti G, Lepier A, Sánchez R, Simon-Ebert T, Schroeder T, Götz M and Berninger B
(See online at https://doi.org/10.1038/nprot.2010.188) - (2011). Long-term single-cell imaging of mammalian stem cells. Nature Methods, 8: S30-35
Schroeder T
(See online at https://doi.org/10.1038/nmeth.1577) - (2011). Using an adherent cell culture of the mouse subependymal zone to study the behavior of adult neural stem cells on a single-cell level. Nature Protocols, 6: 1847-1859
Ortega F, Costa MR, Simon-Ebert T, Schroeder T, Götz M and Berninger B
(See online at https://doi.org/10.1038/nprot.2011.404) - (2013). An automatic pipeline for robust cell detection in bright field images of high-throughput microscopy. BMC Bioinformatics, 14: 297
Buggenthin F, Marr C, Schwarzfischer M, Hoppe PS, Hilsenbeck O, Schroeder T and Theis FJ
(See online at https://doi.org/10.1186/1471-2105-14-297) - (2013). Biallelic expression of nanog protein in mouse embryonic stem cells. Cell Stem Cell, 13: 12-31
Filipczyk A, Gkatzis K, Fu J, Hoppe PS, Lickert H, Anastassiadis K and Schroeder T
(See online at https://doi.org/10.1016/j.stem.2013.04.025) - (2013). Oligodendrogliogenic and neurogenic adult subependymal zone neural stem cells constitute distinct lineages and exhibit differential responsiveness to Wnt signalling. Nature Cell Biology, 15: 602-613
Ortega F, Gascón S, Masserdotti G, Deshpande A, Simon C, Fischer J, Dimou L, Lie DC, Schroeder T and Berninger B
(See online at https://doi.org/10.1038/ncb2736)
