Hypoxia facilitates proliferation of smooth muscle cells derived from pluripotent stem cells for vascular tissue engineering

Fang, L; Mei, J; Yao, Boqian; Liu, Jiang; Liu, Peng; Wang, Xichun; Zhou, Jian; Lin, Zhanyi

doi:10.1002/term.3324

Cited by 3 publications

(4 citation statements)

References 53 publications

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“…Moreover, bioreactor-expanded VSMCs contributed to blood vessel formation in vivo, while also retaining similar expression levels of VSMCs markers compared with 2D cultured VSMCs [ 60 ]. Also aiming to improve hiPSC-derived VSMC production, Fang et al proposed a hypoxic (5% O 2 ) treatment during differentiation, effectively inducing proliferative hiPSC-derived VSMCs, via embryoid body-based differentiation [ 61 ]. The hypoxic conditions enhanced the formation, adhesion and amplification rates of embryoid bodies, and upon directed differentiation, hiPSC-VSMCs exhibited increased cell viability compared to culture under atmospheric air [ 61 ].…”

Section: Strategies For Advancing Smc and Skmc Manufacturingmentioning

confidence: 99%

“…Also aiming to improve hiPSC-derived VSMC production, Fang et al proposed a hypoxic (5% O 2 ) treatment during differentiation, effectively inducing proliferative hiPSC-derived VSMCs, via embryoid body-based differentiation [ 61 ]. The hypoxic conditions enhanced the formation, adhesion and amplification rates of embryoid bodies, and upon directed differentiation, hiPSC-VSMCs exhibited increased cell viability compared to culture under atmospheric air [ 61 ]. Envisaging the application of cell or tissue-based products in clinical practice, it is imperative to adapt protocols towards xeno(geneic)-free conditions, since animal-derived reagents may carry zoonoses and trigger immune responses of cell or tissue derivatives, which could lead to graft failure, besides being associated with batch-to-batch variability and lack of standardization [ 62 ].…”

Section: Strategies For Advancing Smc and Skmc Manufacturingmentioning

confidence: 99%

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Bioprocessing Considerations towards the Manufacturing of Therapeutic Skeletal and Smooth Muscle Cells

Franchi-Mendes,

Silva,

Cartaxo

et al. 2023

Bioengineering

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Tissue engineering approaches within the muscle context represent a promising emerging field to address the current therapeutic challenges related with multiple pathological conditions affecting the muscle compartments, either skeletal muscle or smooth muscle, responsible for involuntary and voluntary contraction, respectively. In this review, several features and parameters involved in the bioprocessing of muscle cells are addressed. The cell isolation process is depicted, depending on the type of tissue (smooth or skeletal muscle), followed by the description of the challenges involving the use of adult donor tissue and the strategies to overcome the hurdles of reaching relevant cell numbers towards a clinical application. Specifically, the use of stem/progenitor cells is highlighted as a source for smooth and skeletal muscle cells towards the development of a cellular product able to maintain the target cell’s identity and functionality. Moreover, taking into account the need for a robust and cost-effective bioprocess for cell manufacturing, the combination of muscle cells with biomaterials and the need for scale-up envisioning clinical applications are also approached.

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Section: Strategies For Advancing Smc and Skmc Manufacturingmentioning

confidence: 99%

Section: Strategies For Advancing Smc and Skmc Manufacturingmentioning

confidence: 99%

Bioprocessing Considerations towards the Manufacturing of Therapeutic Skeletal and Smooth Muscle Cells

Franchi-Mendes,

Silva,

Cartaxo

et al. 2023

Bioengineering

View full text Add to dashboard Cite

show abstract

“…To address these de ciencies, numerous studies have been conducted to create bionic microenvironments that regulate the cells phenotypic behavior and function. These studies have explored various aspects, including threedimensional culture (8), mechanical cues (9), oxygen concentration (10), growth factors (11), and surface morphology (12). However, no studies have been conducted to regulate the phenotypic behavior and function of VSMCs by macromolecular crowding (MMC).…”

Section: Introductionmentioning

confidence: 99%

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing Macromolecular crowding in vitro

Liu,

Jiang,

et al. 2024

Preprint

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Background The contractile phenotype of vascular smooth muscle cells (VSMCs) has a good diastolic and contractile capacity, and their altered function is the main pathophysiological basis for diseases such as hypertension. VSMCs always exist in a synthetic phenotype in vitro, making it challenging to maintain the contractile phenotype for research. It is widely recognized that common medium in vitro is significantly less crowded than the in vivo environment. Additionally, VSMCs have a heightened sense of touch to detect medium crowding changes. However, it is unclear whether macromolecular crowding (MMC) can help maintain the contractile phenotype of VSMCs. Purpose To study and obtain the phenotypic, behavioral and gene expression changes of VSMCs after increasing the crowding degree by adding carrageenan (CR). Methods The medium crowding degree was examined by the dynamic light scattering assay;VSMCs survival and activity were examined by the Calcein/PI cell activity and toxicity assay and CCK-8 assays༛VSMCs phenotype and migration were examined by WB and wound healing assay༛Gene expression was examined by transcriptomic analysis and RT-qPCR. Results 225 µg/mL CR significantly increased the crowding degree of the medium and did not affect cell survival. Simultaneously, CR significantly promoted contraction phenotypic markers expression in VSMCs, cell length was shortened, cell proliferation was decreased, and cell migration was inhibited. CR significantly altered gene expression in VSMCs. Specifically, 856 genes were up-regulated and 1207 genes were down-regulated. These alterations primarily affected cellular ion channel transport, microtubule movement, cellular respiratory metabolism, amino acid transport, and extracellular matrix synthesis. The up-regulated genes were primarily involved in the cytoskeleton and the contraction process of VSMCs, while the down-regulated genes were mainly involved in extracellular matrix synthesis. Conclusion The study showed that VSMCs in vitro can maintain the contractile phenotype by sensing changes in the culture environment's crowding, which can be maintained by adding a concentration of CR.

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“…To address these deficiencies, numerous studies have been conducted to create bionic microenvironments that regulate cellular phenotypic behavior and function. These studies have explored various aspects, including three-dimensional culture [ 6 ], mechanical cues [ 7 ], oxygen concentration [ 8 ], growth factors [ 9 ], and surface morphology [ 10 ]. However, the regulation of the VSMCs contractile phenotype, behavior, and function by MMC remains unexplored.…”

Section: Introductionmentioning

confidence: 99%

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing macromolecular crowding in vitro

Liu,

Jiang,

et al. 2024

Eur J Med Res

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Background The contractile phenotype of vascular smooth muscle cells (VSMCs) results in good diastolic and contractile capacities, and its altered function is the main pathophysiological basis for diseases such as hypertension. VSMCs exist as a synthetic phenotype in vitro, making it challenging to maintain a contractile phenotype for research. It is widely recognized that the common medium in vitro is significantly less crowded than in the in vivo environment. Additionally, VSMCs have a heightened sense for detecting changes in medium crowding. However, it is unclear whether macromolecular crowding (MMC) helps maintain the VSMCs contractile phenotype. Purpose This study aimed to explore the phenotypic, behavioral and gene expression changes of VSMCs after increasing the crowding degree by adding carrageenan (CR). Methods The degree of medium crowding was examined by a dynamic light scattering assay; VSMCs survival and activity were examined by calcein/PI cell activity and toxicity and CCK-8 assays; VSMCs phenotypes and migration were examined by WB and wound healing assays; and gene expression was examined by transcriptomic analysis and RT-qPCR. Results Notably, 225 μg/mL CR significantly increased the crowding degree of the medium and did not affect cell survival. Simultaneously, CR significantly promoted the contraction phenotypic marker expression in VSMCs, shortened cell length, decreased cell proliferation, and inhibited cell migration. CR significantly altered gene expression in VSMCs. Specifically, 856 genes were upregulated and 1207 genes were downregulated. These alterations primarily affect the cellular ion channel transport, microtubule movement, respiratory metabolism, amino acid transport, and extracellular matrix synthesis. The upregulated genes were primarily involved in the cytoskeleton and contraction processes of VSMCs, whereas the downregulated genes were mainly involved in extracellular matrix synthesis. Conclusions The in vitro study showed that VSMCs can maintain the contractile phenotype by sensing changes in the crowding of the culture environment, which can be maintained by adding CR. Graphical Abstract

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Hypoxia facilitates proliferation of smooth muscle cells derived from pluripotent stem cells for vascular tissue engineering

Cited by 3 publications

References 53 publications

Bioprocessing Considerations towards the Manufacturing of Therapeutic Skeletal and Smooth Muscle Cells

Bioprocessing Considerations towards the Manufacturing of Therapeutic Skeletal and Smooth Muscle Cells

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing Macromolecular crowding in vitro

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing macromolecular crowding in vitro

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