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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.


Cardiomyocyte (CM) specific gene expression profiles, as determined by RT-PCR and immunostaining, of CM derived from mouse embryonic stem (ES) cells, Nanog induced pluripotent stem (iPS) cells, and Fbx15-iPS cells. (a) CM-associated structural protein gene expression profiles. The expression of Actn1, Myh6, Myh7, Myl7, Myl2, and Nppb was analyzed by semiquantitative RT-PCR analysis. GAPDH was used as an internal control. (b) Immunofluorescent staining of typical CM-specific proteins on day 15 of differentiation in CM derived from ES, Nanog-iPS, and Fbx15-iPS cells. Cells were stained with Nkx2.5 (green), GATA4 (green), atrial natriuretic peptide (ANP; green), myosin heavy chain (MHC; red), and α-actinin (red). Scale bar = 50 μm.
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fig4: Cardiomyocyte (CM) specific gene expression profiles, as determined by RT-PCR and immunostaining, of CM derived from mouse embryonic stem (ES) cells, Nanog induced pluripotent stem (iPS) cells, and Fbx15-iPS cells. (a) CM-associated structural protein gene expression profiles. The expression of Actn1, Myh6, Myh7, Myl7, Myl2, and Nppb was analyzed by semiquantitative RT-PCR analysis. GAPDH was used as an internal control. (b) Immunofluorescent staining of typical CM-specific proteins on day 15 of differentiation in CM derived from ES, Nanog-iPS, and Fbx15-iPS cells. Cells were stained with Nkx2.5 (green), GATA4 (green), atrial natriuretic peptide (ANP; green), myosin heavy chain (MHC; red), and α-actinin (red). Scale bar = 50 μm.

Mentions: Because iPS cell-derived cardiomyocytes continue to express transgenes at various levels, semiquantitative RT-PCR and immunostaining were used to evaluate the expression of cardiac-specific genes in ES, Nanog-iPS, and Fbx15-iPS cell-derived cardiomyocytes. As shown in Figure 4(a), RT-PCR analysis revealed the appropriate expression of various cardiac marker genes, including α myosin heavy chain (Myh6), β myosin heavy chain (Myh7), myosin light chain-2a (Myl7), Myl2, α-sarcomeric actinin (Actn1), and natriuretic peptide type B (Nppb). Immunofluorescence analyses revealed that the ES and iPS cell-derived cardiomyocytes showed typical sarcomere formation based on positive staining with antibodies directed against α-actinin and myosin heavy chain (MHC), proper nuclear expression of the cardiac-specific transcription factors Nkx2.5 and GATA4, and the presence of atrial natriuretic peptide (ANP) in the secretory granules surrounding the nuclei (Figure 4(b)).


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 (CM) specific gene expression profiles, as determined by RT-PCR and immunostaining, of CM derived from mouse embryonic stem (ES) cells, Nanog induced pluripotent stem (iPS) cells, and Fbx15-iPS cells. (a) CM-associated structural protein gene expression profiles. The expression of Actn1, Myh6, Myh7, Myl7, Myl2, and Nppb was analyzed by semiquantitative RT-PCR analysis. GAPDH was used as an internal control. (b) Immunofluorescent staining of typical CM-specific proteins on day 15 of differentiation in CM derived from ES, Nanog-iPS, and Fbx15-iPS cells. Cells were stained with Nkx2.5 (green), GATA4 (green), atrial natriuretic peptide (ANP; green), myosin heavy chain (MHC; red), and α-actinin (red). Scale bar = 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Cardiomyocyte (CM) specific gene expression profiles, as determined by RT-PCR and immunostaining, of CM derived from mouse embryonic stem (ES) cells, Nanog induced pluripotent stem (iPS) cells, and Fbx15-iPS cells. (a) CM-associated structural protein gene expression profiles. The expression of Actn1, Myh6, Myh7, Myl7, Myl2, and Nppb was analyzed by semiquantitative RT-PCR analysis. GAPDH was used as an internal control. (b) Immunofluorescent staining of typical CM-specific proteins on day 15 of differentiation in CM derived from ES, Nanog-iPS, and Fbx15-iPS cells. Cells were stained with Nkx2.5 (green), GATA4 (green), atrial natriuretic peptide (ANP; green), myosin heavy chain (MHC; red), and α-actinin (red). Scale bar = 50 μm.
Mentions: Because iPS cell-derived cardiomyocytes continue to express transgenes at various levels, semiquantitative RT-PCR and immunostaining were used to evaluate the expression of cardiac-specific genes in ES, Nanog-iPS, and Fbx15-iPS cell-derived cardiomyocytes. As shown in Figure 4(a), RT-PCR analysis revealed the appropriate expression of various cardiac marker genes, including α myosin heavy chain (Myh6), β myosin heavy chain (Myh7), myosin light chain-2a (Myl7), Myl2, α-sarcomeric actinin (Actn1), and natriuretic peptide type B (Nppb). Immunofluorescence analyses revealed that the ES and iPS cell-derived cardiomyocytes showed typical sarcomere formation based on positive staining with antibodies directed against α-actinin and myosin heavy chain (MHC), proper nuclear expression of the cardiac-specific transcription factors Nkx2.5 and GATA4, and the presence of atrial natriuretic peptide (ANP) in the secretory granules surrounding the nuclei (Figure 4(b)).

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.