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Distinct iPS Cells Show Different Cardiac Differentiation Efficiency.

Ohno Y, Yuasa S, Egashira T, Seki T, Hashimoto H, Tohyama S, Saito Y, Kunitomi A, Shimoji K, Onizuka T, Kageyama T, Yae K, Tanaka T, Kaneda R, Hattori F, Murata M, Kimura K, Fukuda K - Stem Cells Int (2013)

Bottom Line: Despite the development of various methods for the generation of iPS cells that have resulted in increased efficiency, safety, and general versatility, it remains unknown which types of iPS cells are suitable for clinical use.We found that high-quality iPS cells exhibited better cardiomyocyte differentiation in terms of the time course and efficiency of differentiation than low-quality iPS cells, which hardly ever differentiated into cardiomyocytes.Because of the different properties of the various iPS cell lines such as cardiac differentiation efficiency and potential safety hazards, newly established iPS cell lines must be characterized prior to their use in cardiac regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.

ABSTRACT
Patient-specific induced pluripotent stem (iPS) cells can be generated by introducing transcription factors that are highly expressed in embryonic stem (ES) cells into somatic cells. This opens up new possibilities for cell transplantation-based regenerative medicine by overcoming the ethical issues and immunological problems associated with ES cells. Despite the development of various methods for the generation of iPS cells that have resulted in increased efficiency, safety, and general versatility, it remains unknown which types of iPS cells are suitable for clinical use. Therefore, the aims of the present study were to assess (1) the differentiation potential, time course, and efficiency of different types of iPS cell lines to differentiate into cardiomyocytes in vitro and (2) the properties of the iPS cell-derived cardiomyocytes. We found that high-quality iPS cells exhibited better cardiomyocyte differentiation in terms of the time course and efficiency of differentiation than low-quality iPS cells, which hardly ever differentiated into cardiomyocytes. Because of the different properties of the various iPS cell lines such as cardiac differentiation efficiency and potential safety hazards, newly established iPS cell lines must be characterized prior to their use in cardiac regenerative medicine.

No MeSH data available.


Related in: MedlinePlus

Cardiomyocyte differentiation efficiency of pluripotent stem cells and temporal gene expression patterns during cardiomyocyte differentiation. (a) Percentage of beating colonies on days 6–15 in mouse embryonic stem (ES) cells, Nanog induced pluripotent stem (iPS) cells, and Fbx15-iPS cells. Data are the mean ± SEM (n = 5 in all groups). ((b)–(g)) Quantitative RT-PCR analyses showing temporal gene expression patterns of the mesodermal marker Brachyury T (b), the early cardiac mesodermal marker Mesp1 (c), the cardiac-specific transcription factors Nkx2.5 (d) and Gata4 (e), and the cardiac-specific proteins Nppa (f) and Myl2 (g) in EB3 ES cells (closed squares), 20D17 Nanog-iPS cells (open circles), and WT-1 Fbx15-iPS cells (open triangles). Data are the mean ± SEM (n = 5 in all groups).
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fig2: Cardiomyocyte differentiation efficiency of pluripotent stem cells and temporal gene expression patterns during cardiomyocyte differentiation. (a) Percentage of beating colonies on days 6–15 in mouse embryonic stem (ES) cells, Nanog induced pluripotent stem (iPS) cells, and Fbx15-iPS cells. Data are the mean ± SEM (n = 5 in all groups). ((b)–(g)) Quantitative RT-PCR analyses showing temporal gene expression patterns of the mesodermal marker Brachyury T (b), the early cardiac mesodermal marker Mesp1 (c), the cardiac-specific transcription factors Nkx2.5 (d) and Gata4 (e), and the cardiac-specific proteins Nppa (f) and Myl2 (g) in EB3 ES cells (closed squares), 20D17 Nanog-iPS cells (open circles), and WT-1 Fbx15-iPS cells (open triangles). Data are the mean ± SEM (n = 5 in all groups).

Mentions: To examine differences in the efficiency of cardiac differentiation, the conventional hanging drop method was used to induce spontaneously beating embryoid bodies (EBs), which contain a rich population of cardiomyocytes. Beating efficiency of EBs is well correlated with cardiomyocyte differentiation from ES and iPS cells and can be a good marker for cardiomyocyte differentiation efficiency [5, 21]. Around day 6-7 after EB formation, beating EBs started to be seen in the ES cells and in all Nanog-iPS cell lines. However, it took longer for the Fbx15-iPS cell lines to differentiate into beating EBs (Figure 2(a)). Furthermore, the incidence of beating EBs was significantly lower in the Fbx15-iPS cell lines. Interestingly, the beating efficiency differed between the three Nanog-iPS cell lines evaluated. Specifically, the incidence of beating EBs was significantly higher in the 20D17 and 38D2 Nanog-iPS cell lines than in both the 38C2 Nanog-iPS cell line and the Fbx15-iPS cell lines, although the incidence of beating EBs remained significantly lower than that in ES cells. There were no significant differences between the 38C2 Nanog-iPS cell line and the Fbx15-iPS cell lines after day 12, although there were differences in the time until spontaneous beating was observed in the 38C2 Nanog-iPS cell line and Fbx15-iPS cell lines. These results indicate that there are differences in the efficiency of cardiac differentiation as well as in the time required for cardiac differentiation, between individual iPS cell lines.


Distinct iPS Cells Show Different Cardiac Differentiation Efficiency.

Ohno Y, Yuasa S, Egashira T, Seki T, Hashimoto H, Tohyama S, Saito Y, Kunitomi A, Shimoji K, Onizuka T, Kageyama T, Yae K, Tanaka T, Kaneda R, Hattori F, Murata M, Kimura K, Fukuda K - Stem Cells Int (2013)

Cardiomyocyte differentiation efficiency of pluripotent stem cells and temporal gene expression patterns during cardiomyocyte differentiation. (a) Percentage of beating colonies on days 6–15 in mouse embryonic stem (ES) cells, Nanog induced pluripotent stem (iPS) cells, and Fbx15-iPS cells. Data are the mean ± SEM (n = 5 in all groups). ((b)–(g)) Quantitative RT-PCR analyses showing temporal gene expression patterns of the mesodermal marker Brachyury T (b), the early cardiac mesodermal marker Mesp1 (c), the cardiac-specific transcription factors Nkx2.5 (d) and Gata4 (e), and the cardiac-specific proteins Nppa (f) and Myl2 (g) in EB3 ES cells (closed squares), 20D17 Nanog-iPS cells (open circles), and WT-1 Fbx15-iPS cells (open triangles). Data are the mean ± SEM (n = 5 in all groups).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig2: Cardiomyocyte differentiation efficiency of pluripotent stem cells and temporal gene expression patterns during cardiomyocyte differentiation. (a) Percentage of beating colonies on days 6–15 in mouse embryonic stem (ES) cells, Nanog induced pluripotent stem (iPS) cells, and Fbx15-iPS cells. Data are the mean ± SEM (n = 5 in all groups). ((b)–(g)) Quantitative RT-PCR analyses showing temporal gene expression patterns of the mesodermal marker Brachyury T (b), the early cardiac mesodermal marker Mesp1 (c), the cardiac-specific transcription factors Nkx2.5 (d) and Gata4 (e), and the cardiac-specific proteins Nppa (f) and Myl2 (g) in EB3 ES cells (closed squares), 20D17 Nanog-iPS cells (open circles), and WT-1 Fbx15-iPS cells (open triangles). Data are the mean ± SEM (n = 5 in all groups).
Mentions: To examine differences in the efficiency of cardiac differentiation, the conventional hanging drop method was used to induce spontaneously beating embryoid bodies (EBs), which contain a rich population of cardiomyocytes. Beating efficiency of EBs is well correlated with cardiomyocyte differentiation from ES and iPS cells and can be a good marker for cardiomyocyte differentiation efficiency [5, 21]. Around day 6-7 after EB formation, beating EBs started to be seen in the ES cells and in all Nanog-iPS cell lines. However, it took longer for the Fbx15-iPS cell lines to differentiate into beating EBs (Figure 2(a)). Furthermore, the incidence of beating EBs was significantly lower in the Fbx15-iPS cell lines. Interestingly, the beating efficiency differed between the three Nanog-iPS cell lines evaluated. Specifically, the incidence of beating EBs was significantly higher in the 20D17 and 38D2 Nanog-iPS cell lines than in both the 38C2 Nanog-iPS cell line and the Fbx15-iPS cell lines, although the incidence of beating EBs remained significantly lower than that in ES cells. There were no significant differences between the 38C2 Nanog-iPS cell line and the Fbx15-iPS cell lines after day 12, although there were differences in the time until spontaneous beating was observed in the 38C2 Nanog-iPS cell line and Fbx15-iPS cell lines. These results indicate that there are differences in the efficiency of cardiac differentiation as well as in the time required for cardiac differentiation, between individual iPS cell lines.

Bottom Line: Despite the development of various methods for the generation of iPS cells that have resulted in increased efficiency, safety, and general versatility, it remains unknown which types of iPS cells are suitable for clinical use.We found that high-quality iPS cells exhibited better cardiomyocyte differentiation in terms of the time course and efficiency of differentiation than low-quality iPS cells, which hardly ever differentiated into cardiomyocytes.Because of the different properties of the various iPS cell lines such as cardiac differentiation efficiency and potential safety hazards, newly established iPS cell lines must be characterized prior to their use in cardiac regenerative medicine.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Keio University School of Medicine, Tokyo 160-8582, Japan.

ABSTRACT
Patient-specific induced pluripotent stem (iPS) cells can be generated by introducing transcription factors that are highly expressed in embryonic stem (ES) cells into somatic cells. This opens up new possibilities for cell transplantation-based regenerative medicine by overcoming the ethical issues and immunological problems associated with ES cells. Despite the development of various methods for the generation of iPS cells that have resulted in increased efficiency, safety, and general versatility, it remains unknown which types of iPS cells are suitable for clinical use. Therefore, the aims of the present study were to assess (1) the differentiation potential, time course, and efficiency of different types of iPS cell lines to differentiate into cardiomyocytes in vitro and (2) the properties of the iPS cell-derived cardiomyocytes. We found that high-quality iPS cells exhibited better cardiomyocyte differentiation in terms of the time course and efficiency of differentiation than low-quality iPS cells, which hardly ever differentiated into cardiomyocytes. Because of the different properties of the various iPS cell lines such as cardiac differentiation efficiency and potential safety hazards, newly established iPS cell lines must be characterized prior to their use in cardiac regenerative medicine.

No MeSH data available.


Related in: MedlinePlus