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
. 2022 Oct 11;17(10):2272-2285.
doi: 10.1016/j.stemcr.2022.08.007. Epub 2022 Sep 8.

Space microgravity improves proliferation of human iPSC-derived cardiomyocytes

Antonio Rampoldi  1 , Parvin Forghani  1 , Dong Li  1 , Hyun Hwang  1 , Lawrence Christian Armand  1 , Jordan Fite  2 , Gene Boland  2 , Joshua Maxwell  1 , Kevin Maher  1 , Chunhui Xu  3
Affiliations

Space microgravity improves proliferation of human iPSC-derived cardiomyocytes

Antonio Rampoldi et al. Stem Cell Reports. .

Abstract

In microgravity, cells undergo profound changes in their properties. However, how human cardiac progenitors respond to space microgravity is unknown. In this study, we evaluated the effect of space microgravity on differentiation of human induced pluripotent stem cell (hiPSC)-derived cardiac progenitors compared with 1G cultures on the International Space Station (ISS). Cryopreserved 3D cardiac progenitors were cultured for 3 weeks on the ISS. Compared with 1G cultures, the microgravity cultures had 3-fold larger sphere sizes, 20-fold higher counts of nuclei, and increased expression of proliferation markers. Highly enriched cardiomyocytes generated in space microgravity showed improved Ca2+ handling and increased expression of contraction-associated genes. Short-term exposure (3 days) of cardiac progenitors to space microgravity upregulated genes involved in cell proliferation, survival, cardiac differentiation, and contraction, consistent with improved microgravity cultures at the late stage. These results indicate that space microgravity increased proliferation of hiPSC-cardiomyocytes, which had appropriate structure and function.

Keywords: calcium handling; cardiac progenitors; cardiomyocytes; differentiation; function; gene experssison; human induced pluripotent stem cells; microgravity; proliferation; spaceflight.

PubMed Disclaimer

Conflict of interest statement

Conflicts of interest J.F. and G.B. were employees of Techshot. All other authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Spaceflight experimental design (A) Schematic of spaceflight operational plan. Scale bar, 200 μm. (B) Schematic of Multi-specimen Variable-gravity Platform (MVP) sample collection plan. (C) Schematic of MVP system. ICC, immunocytochemistry. See also Figure S1.
Figure 2
Figure 2
Space microgravity increases cardiac sphere size and improves proliferation of enriched hiPSC-CMs (A) Cell morphology of cardiac spheres derived from SCVI-273 and IMR90 hiPSCs from International Space Station (ISS) cultures post flight (scale bar, 200 μm). (B) Diameters of cardiac spheres derived from SCVI-273 and IMR90 hiPSCs (n = 72–180 spheres). (C) Immunocytochemistry analysis of IMR90 cultures to detect cardiomyocyte purity by cardiac marker NKX2.5 (red) and proliferation by Ki-67 (green) and cardiac marker cTNI (red) (scale bar, 50μm). (D) Percentage of NKX2.5-positive cells and Ki-67-positive cells in IMR90 cultures. Aliquots of ISS cultures post flight were dissociated, replated, and subjected to immunocytochemistry (n = 3 cultures). (E) Expression of genes associated with proliferation in IMR90 cultures (3 weeks on the ISS) and SCVI-273 cultures (3 days on the ISS) (n = 3 cultures). (F) Counts of nuclei in samples from IMR90 cultures (n = 6 wells) after the cells were dissociated, replated, and stained. Statistical analyses were performed with unpaired, two-tailed Student’s t test for (B), (D), and (F), and one-way ANOVA for (E). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. Data are presented as mean ± SD. ISS 1G, the 1G condition on the ISS; ISS μG, the microgravity condition on the ISS.
Figure 3
Figure 3
Space microgravity cultures generated enriched cardiomyocytes with increased cell size (A) Immunocytochemical analysis of cardiac proteins in IMR90 cells from ISS cultures post flight, including cTNT (red), pan-cadherin (red), α-actinin (green), and NKX2.5 (red) (scale bar, 50 μm). (B) Comparison of cellular parameters of IMR90 hiPSC-CMs. Statistical analyses were performed with unpaired, two-tailed Student’s t test. ∗∗p < 0.01, and ∗∗∗∗p < 0.0001. Data are presented as mean ± SD (ISS 1G, n = 53 cells; ISS μG, n = 73 cells).
Figure 4
Figure 4
Space microgravity improves cardiac structure of hiPSC-CMs (A) Structural analysis of IMR90 hiPSC-CMs from ISS cultures post flight. Cells were dissociated, replated, and stained for sarcomeric α-actinin (green) and Hoechst (blue). Overall appearance of myofibrillar structure was categorized into three different levels and percentage of the cells by the scores was generated by counting n = 53 cells from ISS 1G and n = 73 cells from ISS μG cultures. (B) qRT-PCR panel showing relative mRNA expression levels of gene associated with cardiac structure in cells derived from IMR90 hiPSCs (3 weeks on the ISS) and SCVI-273 hiPSCs (3 days on the ISS) (n = 3 cultures). Statistical analyses was performed with one-way ANOVA. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. Data are presented as mean ± SD (n = 3 cultures).
Figure 5
Figure 5
Space microgravity improves intracellular Ca2+ handling in hiPSC-CMs (A) Representative traces of normal Ca2+ transients or abnormal Ca2+ transients of IMR90 hiPSC-CMs from ISS cultures post flight. (B) Pie chart showing the percentage of cells exhibiting normal Ca2+ transients or abnormal Ca2+ transients (ISS 1G, n = 36 cells; ISS μG, n = 81 cells). (C) Ca2+ transient analyses with parameters presented as mean ± SD (ISS 1G, n = 28 cells; ISS μG, n = 75 cells). (D) qRT-PCR panel showing relative mRNA expression levels of genes associated with Ca2+ handling in cells derived from IMR90 hiPSCs (3 weeks on the ISS) and SCVI-273 hiPSCs (3 days on the ISS) (n = 3 cultures). Statistical analyses were performed with unpaired, two-tailed Student’s t test for (C) and one-way ANOVA for (D). ∗∗∗p < 0.001; and ∗∗∗∗p < 0.0001.
Figure 6
Figure 6
Differentially expressed genes and GO terms identified by RNA sequencing analysis of hiPSC-CMs exposed to space microgravity (A) Volcano plot illustrating differentially expressed genes between ISS μG and ISS 1G samples (n = 3 cultures) collected from SCVI-273 cultures of 3 days on the ISS (short-term exposure to microgravity). (B) Dot plot showing up- and downregulated GO terms of biological processes. GSEA, Gene Set Enrichment Analysis. See also Figures S2–S7.
Figure 7
Figure 7
Chord diagrams showing the relationship between GO terms and differentially expressed genes in hiPSC-CMs exposed to space microgravity (A) Chord diagram of selected upregulated GO terms and genes in ISS μG versus ISS 1G of SCVI-273 cultures of 3 days on the ISS. (B) Chord diagram of selected downregulated GO terms and genes. GO terms are presented on the right, genes on the left, and colored squares on the left indicate Log2(fold change) value from highest to lowest (n = 3 cultures).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Baio J., Martinez A.F., Bailey L., Hasaniya N., Pecaut M.J., Kearns-Jonker M. Spaceflight activates protein kinase C alpha signaling and modifies the developmental stage of human neonatal cardiovascular progenitor cells. Stem Cells Dev. 2018;27:805–818. doi: 10.1089/scd.2017.0263. - DOI - PubMed
    1. Baio J., Martinez A.F., Silva I., Hoehn C.V., Countryman S., Bailey L., Hasaniya N., Pecaut M.J., Kearns-Jonker M. Cardiovascular progenitor cells cultured aboard the International Space Station exhibit altered developmental and functional properties. NPJ Microgravity. 2018;4:13. doi: 10.1038/s41526-018-0048-x. - DOI - PMC - PubMed
    1. Baljinnyam E., Venkatesh S., Gordan R., Mareedu S., Zhang J., Xie L.H., Azzam E.I., Suzuki C.K., Fraidenraich D. Effect of densely ionizing radiation on cardiomyocyte differentiation from human-induced pluripotent stem cells. Physiol. Rep. 2017;5:e13308. doi: 10.14814/phy2.13308. - DOI - PMC - PubMed
    1. Barzegari A., Saei A.A. An update to space biomedical research: tissue engineering in microgravity bioreactors. Bioimpacts. 2012;2:23–32. doi: 10.5681/bi.2012.003. - DOI - PMC - PubMed
    1. Basisty N., Kale A., Jeon O.H., Kuehnemann C., Payne T., Rao C., Holtz A., Shah S., Sharma V., Ferrucci L., et al. A proteomic atlas of senescence-associated secretomes for aging biomarker development. PLoS Biol. 2020;18:e3000599. doi: 10.1371/journal.pbio.3000599. - DOI - PMC - PubMed

Publication types