ES to cardiomyocyte

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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13328-143B7	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day01	biol_rep1
13329-143B8	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day02	biol_rep1
13330-143B9	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day03	biol_rep1
13331-143C1	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day04	biol_rep1
13332-143C2	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day05	biol_rep1
13333-143C3	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day06	biol_rep1
13335-143C5	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day08	biol_rep1
13336-143C6	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day09	biol_rep1
13337-143C7	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day10	biol_rep1
13338-143C8	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day11	biol_rep1
13339-143C9	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day12	biol_rep1
13340-143D1	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day01	biol_rep2
13341-143D2	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day02	biol_rep2
13342-143D3	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day03	biol_rep2
13343-143D4	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day04	biol_rep2
13344-143D5	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day05	biol_rep2
13345-143D6	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day06	biol_rep2
13346-143D7	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day07	biol_rep2
13347-143D8	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day08	biol_rep2
13348-143D9	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day09	biol_rep2
13349-143E1	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day10	biol_rep2
13350-143E2	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day11	biol_rep2
13351-143E3	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day12	biol_rep2
13352-143E4	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day01	biol_rep3
13353-143E5	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day02	biol_rep3
13354-143E6	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day03	biol_rep3
13355-143E7	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day04	biol_rep3
13356-143E8	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day05	biol_rep3
13357-143E9	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day06	biol_rep3
13358-143F1	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day07	biol_rep3
13359-143F2	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day08	biol_rep3
13360-143F3	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day09	biol_rep3
13361-143F4	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day10	biol_rep3
13362-143F5	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day11	biol_rep3
13363-143F6	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day12	biol_rep3
13364-143F7	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day00	biol_rep1 (UH-1)
13365-143F8	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day00	biol_rep2 (UH-2)
13366-143F9	HES3-GFP Embryonic Stem cells, cardiomyocytic induction	day00	biol_rep3 (UH-3)