Enhanced cardiomyogenesis of human embryonic stem cells by a small molecular inhibitor of p38 MAPK
| Autor: | Robert Zweigerdt, Set Yen Soo, Ralph Eberhard Graichen, Christine L. Mummery, Su Chin Tham, Thavamalar Balakrishnan, Siti Norfiza, Alan Colman, Bruce Paul Davidson, Xiuqin Xu, Shirly Sieh, Stefan R. Braam |
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| Přispěvatelé: | Hubrecht Institute for Developmental Biology and Stem Cell Research |
| Rok vydání: | 2008 |
| Předmět: |
Cancer Research
Patch-Clamp Techniques Pyridines Cellular differentiation Population Embryoid body Biology p38 Mitogen-Activated Protein Kinases Culture Media Serum-Free Cell Line Humans education Protein Kinase Inhibitors Molecular Biology Embryonic Stem Cells education.field_of_study Reverse Transcriptase Polymerase Chain Reaction Imidazoles Cell Differentiation Heart Cell Biology Embryonic stem cell Molecular biology In vitro Cell culture Mitogen-activated protein kinase embryonic structures Mesoderm formation biology.protein biological phenomena cell phenomena and immunity Signal Transduction Developmental Biology |
| Zdroj: | Differentiation, 76(4), 357-370. Elsevier B.V. |
| ISSN: | 0301-4681 |
| Popis: | Human embryonic stem cells (hESC) can differentiate to cardiomyocytes in vitro but with generally poor efficiency. Here, we describe a novel method for the efficient generation of cardiomyocytes from hESC in a scalable suspension culture process. Differentiation in serum-free medium conditioned by the cell line END2 (END2-CM) readily resulted in differentiated cell populations with more than 10% cardiomyocytes without further enrichment. By screening candidate molecules, we have identified SB203580, a specific p38 MAP kinase inhibitor, as a potent promoter of hESC-cardiogenesis. SB203580 at concentrations10 microM, induced more than 20% of differentiated cells to become cardiomyocytes and increased total cell numbers, so that the overall cardiomyocyte yield was approximately 2.5-fold higher than controls. Gene expression indicated that early mesoderm formation was favored in the presence of SB203580. Accordingly, transient addition of the inhibitor at the onset of differentiation only was sufficient to determine the hESC fate. Patch clamp electrophysiology showed that the distribution of cardiomyocyte phenotypes in the population was unchanged by the compound. Interestingly, cardiomyogenesis was strongly inhibited at SB203580 concentrationsor =15 microM. Thus, modulation of the p38MAP kinase pathway, in combination with factors released by END2 cells, plays an essential role in early lineage determination in hESC and the efficiency of cardiomyogenesis. Our findings contribute to transforming human cardiomyocyte generation from hESC into a robust and scalable process. |
| Databáze: | OpenAIRE |
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