Stiffness‐Tunable Hydrogel‐Sandwich Culture Modulates the YAP‐Mediated Mechanoresponse in Induced‐Pluripotent Stem Cell Embryoid Bodies and Augments Cardiomyocyte Differentiation

Nattasit, Praphawi; Niibe, Kunimichi; Yamada, Masahiro; Ohori-Morita, Yumi; Limraksasin, Phoonsuk; Tiskratok, Watcharaphol; Yamamoto, Masaya; Egusa, Hiroshi

doi:10.1002/mabi.202300021

Cited by 4 publications

(3 citation statements)

References 70 publications

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“…From EB4d to NPCs, mitochondrial pH increased and NPCs mitochondrial pH increased to 7.9

0.1. The reason for this was that EB changed from an adherent state to a suspension state on day 4, which restricted cell proliferation and increased cell differentiation [ 38 ], as shown in Fig. 3(a3) .…”

Section: Resultsmentioning

confidence: 99%

SERS-based long-term mitochondrial pH monitoring during differentiation of human induced pluripotent stem cells to neural progenitor cells

Yang,

Bu,

et al. 2024

Biomed. Opt. Express

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As one of the important organelles in the process of cell differentiation, mitochondria regulate the whole process of differentiation by participating in energy supply and information transmission. Mitochondrial pH value is a key indicator of mitochondrial function. Therefore, real-time monitoring of mitochondrial pH value during cell differentiation is of great significance for understanding cell biochemical processes and exploring differentiation mechanisms. In this study, Surface-enhanced Raman scattering (SERS) technology was used to achieve the real-time monitoring of mitochondrial pH during induced pluripotent stem cells (iPSCs) differentiation into neural progenitor cells (NPCs). The results showed that the variation trend of mitochondrial pH in normal and abnormal differentiated batches was different. The mitochondrial pH value of normal differentiated cells continued to decline from iPSCs to embryoid bodies (EB) day 4, and continued to rise from EB day 4 to the NPCs stage, and the mitochondrial microenvironment of iPSCs to NPCs differentiation became acidic. In contrast, the mitochondrial pH value of abnormally differentiated cells declined continuously during differentiation. This study improves the information on acid-base balance during cell differentiation and may provide a basis for further understanding of the changes and regulatory mechanisms of mitochondrial metabolism during cell differentiation. This also helps to improve more accurate and useful differentiation protocols based on the microenvironment within the mitochondria, improving the efficiency of cell differentiation.

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“…From EB4d to NPCs, mitochondrial pH increased and NPCs mitochondrial pH increased to 7.9

Section: Resultsmentioning

confidence: 99%

SERS-based long-term mitochondrial pH monitoring during differentiation of human induced pluripotent stem cells to neural progenitor cells

Yang,

Bu,

et al. 2024

Biomed. Opt. Express

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“…In addition, it is necessary to verify whether the stiffness suitable for controlling a certain cell function is universal for other materials or extracellular matrices used in microphysiological systems. For instance, hydrogel-based materials can provide a space in which cell functions can be tuned synergistically by both the static mechanical stimulation of material stiffness and the ligand stimulation of immobilized biomolecule [57]. Further research interests remain in the future.…”

Section: Discussionmentioning

confidence: 99%

Mechanoregulation of Osteoclastogenesis-Inducing Potentials of Fibrosarcoma Cell Line by Substrate Stiffness

Tiskratok,

Yamada,

Watanabe

et al. 2023

IJMS

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A micro-physiological system is generally fabricated using soft materials, such as polydimethylsiloxane silicone (PDMS), and seeks an inflammatory osteolysis model for osteoimmunological research as one of the development needs. Microenvironmental stiffness regulates various cellular functions via mechanotransduction. Controlling culture substrate stiffness may help spatially coordinate the supply of osteoclastogenesis-inducing factors from immortalized cell lines, such as mouse fibrosarcoma L929 cells, within the system. Herein, we aimed to determine the effects of substrate stiffness on the osteoclastogenesis-inducing potential of L929 cells via cellular mechanotransduction. L929 cells showed increased expression of osteoclastogenesis-inducing factors when cultured on type I collagen-coated PDMS substrates with soft stiffness, approximating that of soft tissue sarcomas, regardless of the addition of lipopolysaccharide to augment proinflammatory reactions. Supernatants of L929 cells cultured on soft PDMS substrates promoted osteoclast differentiation of the mouse osteoclast precursor RAW 264.7 by stimulating the expression of osteoclastogenesis-related gene markers and tartrate-resistant acid phosphatase activity. The soft PDMS substrate inhibited the nuclear translocation of YES-associated proteins in L929 cells without reducing cell attachment. However, the hard PDMS substrate hardly affected the cellular response of the L929 cells. Our results showed that PDMS substrate stiffness tuned the osteoclastogenesis-inducing potential of L929 cells via cellular mechanotransduction.

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“…Simple and utilizing novel techniques are expected using the features of 2D and 3D cell culture systems. Sandwich culture is one of the interesting culture systems that cultivate cells on the top and bottom substrates. − The substrates are available as temporal packings and are easy to isolate. Additionally, a variety of 2D substrates with different stiffnesses or alignments can be selected and utilized for precise control.…”

Section: Introductionmentioning

confidence: 99%

Adhesive Sulfabetaine Polymer Hydrogels for the Sandwich Cell Culture

Morimoto,

Murata,

Yamamoto

et al. 2024

ACS Omega

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Sandwich culture systems are techniques that cultivate cells by sandwiching them between the top and bottom substrates. Since the substrates can be separated, the system is expected to be applied to the construct layering of patterned cells and to the isolation of stacked cells. In this study, we prepared hydrogels composed of zwitterionic sulfabetaine polymers, poly[2-(2-(methacryloyloxyethyl)dimethylammonio)ethyl-1-sulfate] (PZBMA). The ZBMA homopolymers have been shown to form aggregates in aqueous solutions due to their intermolecular interactions. The water content of the PZBMA hydrogels in water was ∼70% regardless of N,N′-methylenebis(acrylamide), BIS, content as the cross-linker. The results indicated that the intermolecular interaction contributed more to the swelling behaviors than the chemical cross-linker. However, PZBMA hydrogels with 0.1 mol % BIS showed not only high elongation (∼850%) properties but also high adhesiveness and self-healing properties. When this PZBMA hydrogel was impregnated with collagen and subjected to sandwich culture using Madin−Darby canine kidney (MDCK) cells, a three-dimensional morphology of MDCK cell aggregates was constructed. Such a sulfabetaine hydrogel is expected to be developed for regenerative medicine.

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Stiffness‐Tunable Hydrogel‐Sandwich Culture Modulates the YAP‐Mediated Mechanoresponse in Induced‐Pluripotent Stem Cell Embryoid Bodies and Augments Cardiomyocyte Differentiation

Cited by 4 publications

References 70 publications

SERS-based long-term mitochondrial pH monitoring during differentiation of human induced pluripotent stem cells to neural progenitor cells

SERS-based long-term mitochondrial pH monitoring during differentiation of human induced pluripotent stem cells to neural progenitor cells

Mechanoregulation of Osteoclastogenesis-Inducing Potentials of Fibrosarcoma Cell Line by Substrate Stiffness

Adhesive Sulfabetaine Polymer Hydrogels for the Sandwich Cell Culture

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