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|TCSample_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).<br><br>References:<br>[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.<br>[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.<br><br><br>
|TCSample_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).<br><br>References:<br>[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.<br>[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.<br><br><br>
|Time_Course=
|Time_Course=
|category_treatment=differentiation
|category_treatment=Differentiation
|collaborators=Christine Mummery
|collaborators=Christine Mummery
|description=human_HES3-GFP_Embryonic_Stem_cells
|description=human_HES3-GFP_Embryonic_Stem_cells
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|time_points=day00
|time_points=day00
|time_span=12 days
|time_span=12 days
|timepoint_design=staged in-vitro diff
|timepoint_design=Staged in-vitro diff
|tissue_cell_type=ES->cardiomyocyte
|tissue_cell_type=ES>>cardiomyocyte
|zenbu_config=http://fantom.gsc.riken.jp/zenbu/gLyphs/#config=9aiqvfLkCFxuK8-E4coyRDHuman H3 embryonic stem cells differentiated to cardiomyocytes
|zenbu_config=http://fantom.gsc.riken.jp/zenbu/gLyphs/#config=9aiqvfLkCFxuK8-E4coyRDHuman H3 embryonic stem cells differentiated to cardiomyocytes
}}
}}

Revision as of 12:08, 4 February 2015

Series:IN_VITRO DIFFERENTIATION SERIES
Species:Human (Homo sapiens)
Genomic View:H3 embryonic stem cells differentiated to cardiomyocytes Zenbu
Expression table:FILE
Link to TET:[{{{tet_file}}} 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


13328-143B7HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday01biol_rep1
13329-143B8HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday02biol_rep1
13330-143B9HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday03biol_rep1
13331-143C1HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday04biol_rep1
13332-143C2HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday05biol_rep1
13333-143C3HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday06biol_rep1
13335-143C5HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday08biol_rep1
13336-143C6HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday09biol_rep1
13337-143C7HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday10biol_rep1
13338-143C8HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday11biol_rep1
13339-143C9HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday12biol_rep1
13340-143D1HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday01biol_rep2
13341-143D2HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday02biol_rep2
13342-143D3HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday03biol_rep2
13343-143D4HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday04biol_rep2
13344-143D5HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday05biol_rep2
13345-143D6HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday06biol_rep2
13346-143D7HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday07biol_rep2
13347-143D8HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday08biol_rep2
13348-143D9HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday09biol_rep2
13349-143E1HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday10biol_rep2
13350-143E2HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday11biol_rep2
13351-143E3HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday12biol_rep2
13352-143E4HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday01biol_rep3
13353-143E5HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday02biol_rep3
13354-143E6HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday03biol_rep3
13355-143E7HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday04biol_rep3
13356-143E8HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday05biol_rep3
13357-143E9HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday06biol_rep3
13358-143F1HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday07biol_rep3
13359-143F2HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday08biol_rep3
13360-143F3HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday09biol_rep3
13361-143F4HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday10biol_rep3
13362-143F5HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday11biol_rep3
13363-143F6HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday12biol_rep3
13364-143F7HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday00biol_rep1 (UH-1)
13365-143F8HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday00biol_rep2 (UH-2)
13366-143F9HES3-GFP Embryonic Stem cells, cardiomyocytic inductionday00biol_rep3 (UH-3)
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