Multiplexing physical stimulation on single human induced pluripotent stem cell-derived cardiomyocytes for phenotype modulation

Kit‐Anan, Worrapong; Mazo, Manuel; Wang, Brian; Leonardo, Vincent; Pence, Isaac J.; Gopal, Sahana; Gelmi, Amy; Nagelkerke, Anika; Becce, Michele; Chiappini, Ciro; Harding, Siân E.; Terracciano, Cesare M.; Stevens, Molly M.

doi:10.1088/1758-5090/abce0a

Cited by 13 publications

(12 citation statements)

References 94 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…When interacting with materials, adhered cells are stimulated by various physicochemical signals, including topographical cues, heat, electricity, and magnetism. Among these physical signals, the regulatory effect of substrate physicochemical signals on hPSCs is of most interest to researchers because it is a one-step method that is feasible in most laboratories. , Various projects have illuminated the promoting effects of surface topographical cues such as micro/nanogroove, , pillar, and hierarchical patterns on cardiac differentiation and maturation of hPSCs.…”

Section: Results and Discussionmentioning

confidence: 99%

Binary Colloidal Crystals Promote Cardiac Differentiation of Human Pluripotent Stem Cells via Nuclear Accumulation of SETDB1

et al. 2023

View full text Add to dashboard Cite

Biophysical cues can facilitate the cardiac differentiation of human pluripotent stem cells (hPSCs), yet the mechanism is far from established. One of the binary colloidal crystals, composed of 5 μm Si and 400 nm poly(methyl methacrylate) particles named 5PM, has been applied as a substrate for hPSCs cultivation and cardiac differentiation. In this study, cell nucleus, cytoskeleton, and epigenetic states of human induced pluripotent stem cells on the 5PM were analyzed using atomic force microscopy, molecular biology assays, and the assay for transposase-accessible chromatin sequencing (ATAC-seq). Cells were more spherical with stiffer cell nuclei on the 5PM compared to the flat control. ATAC-seq revealed that chromatin accessibility decreased on the 5PM, caused by the increased entry of histone lysine methyltransferase SETDB1 into the cell nuclei and the amplified level of histone H3K9me3 modification. Reducing cytoskeleton tension using a ROCK inhibitor attenuated the nuclear accumulation of SETDB1 on the 5PM, indicating that the effect is cytoskeleton-dependent. In addition, the knockdown of SETDB1 reversed the promotive effects of the 5PM on cardiac differentiation, demonstrating that biophysical cue-induced cytoskeletal tension, cell nucleus deformation, and then SETDB1 accumulation are critical outside-in signal transformations in cardiac differentiation. Human embryonic stem cells showed similar results, indicating that the biophysical impact of the 5PM surfaces on cardiac differentiation could be universal. These findings contribute to our understanding of material-assistant hPSC differentiation, which benefits materiobiology and stem cell bioengineering.

show abstract

Section: Results and Discussionmentioning

confidence: 99%

Binary Colloidal Crystals Promote Cardiac Differentiation of Human Pluripotent Stem Cells via Nuclear Accumulation of SETDB1

et al. 2023

View full text Add to dashboard Cite

show abstract

“…Furthermore, cell length/width ratio and circularity are also recognized to be valuable analytical tools to determine the structural development and maturation of in vitro hiPSC-CMs . It has been reported that the phenotype of human adult CMs exhibits a length-to-width ratio of around 5–7 . To be able to compare the cell shape on different substrates with adult CM values, we calculated the length/width ratio, that is, the major axis of the cell divided by the minor axis.…”

Section: Results and Discussionmentioning

confidence: 99%

Aligned Nanofibrous Net Deposited Perpendicularly on Microridges Supports Endothelium Formation and Promotes the Structural Maturation of hiPSC-Derived Cardiomyocytes

Tian

Liang

Lin

et al. 2023

ACS Appl. Mater. Interfaces

View full text Add to dashboard Cite

Cell alignment widely exists in various in vivo tissues and also plays an essential role in the construction of in vitro models, such as vascular endothelial and myocardial models. Recently, microscale and nanoscale hierarchical topographical structures have been drawing increasing attention for engineering in vitro cell alignment. In the present study, we fabricated a micro-/nanohierarchical substrate based on soft lithography and electrospinning to assess the synergetic effect of both the aligned nanofibrous topographical guidance and the off-ground culture environment provided by the substrate on the endothelium formation and the maturation of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The morphology, proliferation, and barrier formation of human umbilical vein endothelial cells (HUVECs) as well as the alignment, cardiac-specific proteins, and maturity-related gene expression of hiPSC-CMs on the aligned-nanofiber/microridge (AN-MR) substrate were studied. Compared with the glass slide and the single-aligned nanofiber substrate, the AN-MR substrate enhanced the proliferation, alignment, and cell−cell interaction of HUVECs and improved the length of the sarcomere and maturation-related gene expression of hiPSC-CMs. Finally, the response of hiPSC-CMs on different substrates to two typical cardiac drugs (isoproterenol and E-4031) was tested and analyzed, showing that the hiPSC-CMs on AN-MR substrates were more resistant to drugs than those in other groups, which was related to the higher maturity of the cells. Overall, the proposed micro-/nanohierarchical substrate supports the in vitro endothelium formation and enhances the maturation of hiPSC-CMs, which show great potential to be applied in the construction of in vitro models and tissue engineering.

show abstract

“…In the presented form, the tool of reshaping is not suited for in vivo experimentation, but importantly, the gain of knowledge that both longitudinal shape and t-tubule formation are required for functional maturation, will have to be considered for the preparation of cell grafts. In future studies, the impact of the composition of the extracellular matrix (ECM) and its stiffness [ 25 ] as well as the effect of outside-in signalling [ 23 ] on the functional maturation of these cells must be taken into account. More physiological growth surfaces may also permit prolonged culture times and, thus, a more effective functional maturation [ 48 ] of these cells.…”

Section: Limitations Of the Studymentioning

confidence: 99%

“…Recent research has focussed intensively on the investigation of the effect of topographical and physical cues on the maturation of hiPSC-CM during culture. Cardiomyocytes grown on micropatterned substrates enhanced cell alignment and improved Ca 2+ handling and contraction [ 23 , 35 , 40 , 54 ]. In a recent study, we reported that single mouse iPSC-CM seeded into cuboid 3D micro-scaffolds produced strong tubular invaginations and revealed more robust Ca 2+ signalling [ 46 ].…”

Section: Introductionmentioning

confidence: 99%

Membrane remodelling triggers maturation of excitation–contraction coupling in 3D-shaped human-induced pluripotent stem cell-derived cardiomyocytes

et al. 2023

View full text Add to dashboard Cite

The prospective use of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) for cardiac regenerative medicine strongly depends on the electro-mechanical properties of these cells, especially regarding the Ca2+-dependent excitation–contraction (EC) coupling mechanism. Currently, the immature structural and functional features of hiPSC-CM limit the progression towards clinical applications. Here, we show that a specific microarchitecture is essential for functional maturation of hiPSC-CM. Structural remodelling towards a cuboid cell shape and induction of BIN1, a facilitator of membrane invaginations, lead to transverse (t)-tubule-like structures. This transformation brings two Ca2+ channels critical for EC coupling in close proximity, the L-type Ca2+ channel at the sarcolemma and the ryanodine receptor at the sarcoplasmic reticulum. Consequently, the Ca2+-dependent functional interaction of these channels becomes more efficient, leading to improved spatio-temporal synchronisation of Ca2+ transients and higher EC coupling gain. Thus, functional maturation of hiPSC-cardiomyocytes by optimised cell microarchitecture needs to be considered for future cardiac regenerative approaches.

show abstract

Multiplexing physical stimulation on single human induced pluripotent stem cell-derived cardiomyocytes for phenotype modulation

Abstract: Multiplexing physical stimulation on single human induced pluripotent stem cell-derived cardiomyocytes for phenotype modulation To cite this article: Worrapong Kit-Anan et al 2021 Biofabrication 13 025004 View the article online for updates and enhancements.

Cited by 13 publications

References 94 publications

Binary Colloidal Crystals Promote Cardiac Differentiation of Human Pluripotent Stem Cells via Nuclear Accumulation of SETDB1

Binary Colloidal Crystals Promote Cardiac Differentiation of Human Pluripotent Stem Cells via Nuclear Accumulation of SETDB1

Aligned Nanofibrous Net Deposited Perpendicularly on Microridges Supports Endothelium Formation and Promotes the Structural Maturation of hiPSC-Derived Cardiomyocytes

Membrane remodelling triggers maturation of excitation–contraction coupling in 3D-shaped human-induced pluripotent stem cell-derived cardiomyocytes

Contact Info

Product

Resources

About