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ES to cardiomyocyte

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Series:IN_VITRO DIFFERENTIATION SERIES
Species:Human (Homo sapiens)
Genomic View:Zenbu
Expression table:FILE
Link to TET:TET
Sample providers :Christine Mummery
Germ layer:mesoderm
Primary cells or cell line:cell line
Time span:12 days
Number of time points:13


Overview

Human H3 embryonic stem cells differentiated to cardiomyocytes

Human pluripotent stem cells have the potential to differentiate to all cell types of the human body, including cardiomyocytes. Since the adult human heart does not have the capacity to regenerate, loss of cardiomyocytes after myocardial infarction eventually may lead to heart failure[1]. Cardiomyocytes derived from human pluripotent stem cells may represent a source for future cell replacement in patients with cardiac injury. Furthermore, human cardiomyocytes may also be used for toxicity screening, drug discovery, and for studying mechanisms related to cardiac disease and development. It is important to study the regulatory molecular networks during cardiomyocyte differentiation in order to get a better understanding of processes such as specification, maturation and proliferation of cardiomyocytes. This will lead most likely to novel insights regarding endogenous cardiac regeneration, tissue engineering and cardiac disease.

References:
[1] C. L. Mummery, J. Zhang, E. S. Ng, D. A. Elliott, A. G. Elefanty, and T. J. Kamp, “Differentiation of human embryonic stem cells and induced pluripotent stem cells to cardiomyocytes: a methods overview.,” Circ. Res., vol. 111, no. 3, pp. 344–358, Jul. 2012.

Sample description

For the differentiation of human pluripotent stem cells we used the human embryonic stem cell line HES3-GFP, ubiquitously expressing GFP. Previously, we have shown that co-culture of human embryonic stem cells with a mouse endoderm cell line, END-2, lead to beating cardiomyocytes within 12 days. END-2 cells were treated with mitocmycin C to block proliferation. In general, using this differentiation procedure (in the absence of serum) beating clusters contained 25 % cardiomyocytes[2], [3]. END-2 cells are cultured as a monolayer, whereas differentiated stem cells are grown on top as three-dimensional structures, which allows separation of human embryonic stem cell-derived populations. From undifferentiated human embryonic stem cells and every day during cardiomyocyte differentiation until day 12 samples were collected and used for RNA isolation (n=3).

References:
[2] R. Passier, D. W.-V. Oostwaard, J. Snapper, J. Kloots, R. J. Hassink, E. Kuijk, B. Roelen, A. B. de la Riviere, and C. Mummery, “Increased cardiomyocyte differentiation from human embryonic stem cells in serum-free cultures.,” Stem Cells, vol. 23, no. 6, pp. 772–780, Jun. 2005.
[3] C. Mummery, D. Ward-van Oostwaard, P. Doevendans, R. Spijker, S. van den Brink, R. Hassink, M. van der Heyden, T. Opthof, M. Pera, A. B. de la Riviere, R. Passier, and L. Tertoolen, “Differentiation of human embryonic stem cells to cardiomyocytes: role of coculture with visceral endoderm-like cells.,” Circulation, vol. 107, no. 21, pp. 2733–2740, Jun. 2003.


Quality control



Figure 1: CAGE expression of marker genes in TPM.

References:
[4] A. Beqqali, J. Kloots, D. Ward-van Oostwaard, C. Mummery, and R. Passier, “Genome-wide transcriptional profiling of human embryonic stem cells differentiating to cardiomyocytes.,” Stem Cells, vol. 24, no. 8, pp. 1956–1967, Aug. 2006.

Profiled time course samples

Only samples that passed quality controls (Arner et al. 2015) are shown here. The entire set of samples are downloadable from FANTOM5 human / mouse samples


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