Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2017 Feb 14;8(2):278-289.
doi: 10.1016/j.stemcr.2016.10.009. Epub 2017 Jan 5.

In Vivo Maturation of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Neonatal and Adult Rat Hearts

Shin Kadota  1 , Lil Pabon  1 , Hans Reinecke  1 , Charles E Murry  2
Affiliations

In Vivo Maturation of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Neonatal and Adult Rat Hearts

Shin Kadota et al. Stem Cell Reports. .

Abstract

We hypothesized that the neonatal rat heart would bring transplanted human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to maturity as it grows to adult size. In neonatal rat heart, engrafted hiPSC derivatives developed partially matured myofibrils after 3 months, with increasing cell size and sarcomere length. There was no difference between grafts from hiPSC-CMs or hiPSC-derived cardiac progenitors (hiPSC-CPs) at 3 months, nor was maturation influenced by infarction. Interestingly, the infarcted adult heart induced greater human cardiomyocyte hypertrophy and induction of cardiac troponin I expression than the neonatal heart. Although human cardiomyocytes at all time points were significantly smaller than the host rat cardiomyocytes, transplanted neonatal rat cardiomyocytes reached adult size and structure by 3 months. Thus, the adult rat heart induces faster maturation than the neonatal heart, and human cardiomyocytes mature more slowly than rat cardiomyocytes. The slower maturation of human cardiomyocytes could be related to environmental mismatch or cell-autonomous factors.

Keywords: human pluripotent stem cell-derived cardiomyocyte; maturation; neonatal rat.

PubMed Disclaimer

Figures

None
Graphical abstract
Figure 1
Figure 1
In Vivo Maturation of hiPSC-CMs and Neonatal Rat CMs in Uninjured Growing Rat Hearts (A–F) GFP staining of growing rat hearts at 14 days (A and B), 56 days (C and D), and 84 days (E and F) after hiPSC-CM injection. (G–R) Magnified images of engrafted hiPSC-CMs at 14 days (2w; G–L) and 84 days (3m; M–R) after cell transplantation to neonates. Wheat germ agglutinin (WGA) is used to detect cell membranes, connexin-43 (CX43) is for gap junctions, N-cadherin is for adherence junction, caveolin-3 is for T tubules, α-actinin is for sarcomere, slow skeletal troponin I (ssTNI) is the fetal isoform of TNI, and cardiac TNI (cTNI) is for adult isoform of TNI. (S–X) Magnified images of engrafted NRCMs at 84 days after cell transplantation to neonates. GFP-labeled NRCMs mature as host rat cells. (Y–AA) Comparison of cell diameter (Y), cell sectional area (Z), and sarcomere length (AA) of host rat CMs (n = 34 per group) and hiPSC-CMs (n = 34–40 per group) at 14 days and 84 days after cell injection. At all time points, rat cells are significantly larger than hiPSC-CMs. Data are mean ± SEM. ∗∗p < 0.01 by t test. Scale bars represent 200 μm (A–F) and 10 μm (G–X).
Figure 2
Figure 2
Myocardial Infarction in Neonatal Rat Hearts (A–I) Masson trichrome staining of injured hearts at 3 (A–C), 15 (D–F), and 84 days (G–I) after MI. Scale bars, 1 mm. (J) Scar size showing percentage of left ventricle (%LV) that significantly decreased over time (n = 4). (K) Absolute scar size that significantly increased over time (n = 4). Data are mean ± SEM. ∗∗p < 0.01, ∗p < 0.05 by ANOVA with Bonferroni's post hoc analysis.
Figure 3
Figure 3
In Vivo Maturation of hiPSC-CMs in Injured Growing and Adult Rat Hearts at 3 Months after Cell Injection (A–F) hiPSC-CM injection to neonatal rat MI (A–C) and hiPSC-CM injection to adult rat MI (D–F). Scar tissue is seen as blue by Masson trichrome staining (A and D) and grafts can be seen as brown by GFP staining (B, C, E, and F). (G–R) Magnified images of engrafted hiPSC-CMs at 84 days of transplantation to injured neonatal rat (G–L) and injured adult rat (M–R). Scale bars represent 1 mm (A, B, D, and E), 200 μm (C and F), and 10 μm (G–R). (S–U) Comparison of cell diameter (S), cell sectional area (T), and sarcomere length (U) of hiPSC-CMs (n = 40 per group) engrafted in injured neonatal rat and injured adult rat at 84 days after cell injection. Data are mean ± SEM. ∗∗p < 0.01 by t test.
Figure 4
Figure 4
In Vivo Maturation of hiPSC-Derived Cardiac Progenitors in Uninjured Growing Rat Hearts (A–D) GFP staining of growing rat hearts at 28 days (A and B) and 84 days (C and D) after hiPSC-CP injection. Scale bars, 200 μm. (E–J) Magnified images of engrafted hiPSC-CPs at 84 days of transplantation to uninjured neonatal rats. Scale bars, 10 μm. (K–M) Comparison of cell diameter (K), cell sectional area (L), and sarcomere length (M) of hiPSC-CMs and hiPSC-CPs (n = 40 per group) engrafted in injured neonatal rat at 84 days after cell injection. Data are mean ± SEM. ∗p < 0.05 by t test.
Figure 5
Figure 5
Comparison of hiPSC Derivatives Engrafted in Rat Hearts after 3 Months of Cell Transplantation Cell diameter (A), sarcomere length (B), cell sectional area (C), and myofibril width (D) (n = 40 cells per group). (E) Percentage of cTNI-positive area in grafts (n = 4–5 images of grafts). CP, hiPSC-derived cardiac progenitors; CM, hiPSC-derived cardiomyocytes; neo, neonate. Data are mean ± SEM. ∗∗p < 0.01, ∗p < 0.05 by ANOVA with Bonferroni's post hoc analysis in (C) and (D), and by chi-square test in (E).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Bedada F.B., Chan S.S., Metzger S.K., Zhang L., Zhang J., Garry D.J., Kamp T.J., Kyba M., Metzger J.M. Acquisition of a quantitative, stoichiometrically conserved ratiometric marker of maturation status in stem cell-derived cardiac myocytes. Stem Cell Rep. 2014;3:594–605. - PMC - PubMed
    1. Carson D., Hnilova M., Yang X., Nemeth C.L., Tsui J.H., Smith A.S., Jiao A., Regnier M., Murry C.E., Tamerler C. Nanotopography-induced structural anisotropy and sarcomere development in human cardiomyocytes derived from induced pluripotent stem cells. ACS Appl. Mater. Inter. 2016;8:21923–21932. - PMC - PubMed
    1. Caspi O., Huber I., Kehat I., Habib M., Arbel G., Gepstein A., Yankelson L., Aronson D., Beyar R., Gepstein L. Transplantation of human embryonic stem cell-derived cardiomyocytes improves myocardial performance in infarcted rat hearts. J. Am. Coll. Cardiol. 2007;50:1884–1893. - PubMed
    1. Chong J.J., Yang X., Don C.W., Minami E., Liu Y.W., Weyers J.J., Mahoney W.M., Van Biber B., Cook S.M., Palpant N.J. Human embryonic-stem-cell-derived cardiomyocytes regenerate non-human primate hearts. Nature. 2014;510:273–277. - PMC - PubMed
    1. Denning C., Borgdorff V., Crutchley J., Firth K.S., George V., Kalra S., Kondrashov A., Hoang M.D., Mosqueira D., Patel A. Cardiomyocytes from human pluripotent stem cells: from laboratory curiosity to industrial biomedical platform. Biochim. Biophys. Acta. 2016;1863:1728–1748. - PMC - PubMed

Publication types

LinkOut - more resources

-