Shaking culture enhances chondrogenic differentiation of mouse induced pluripotent stem cell constructs

Limraksasin, Phoonsuk; Kosaka, Yukihiro; Zhang, Maolin; Horie, Naohiro; Kondo, Takeru; Okawa, Hiroko; Yamada, Masahiro; Egusa, Hiroshi

doi:10.1038/s41598-020-72038-y

Cited by 21 publications

(27 citation statements)

References 67 publications

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“…TGF-β signaling is well known to be stimulated in stem cells by mechanical stresses. 10 , 19 Because Tgfb1 was identified as a cell cycle gene differentially expressed by ICF stimulation (Fig. 3b line 3), we focused on TGF-β signaling as a possible mechanism underlying modulation of cell cycle-related gene expression by ICF.…”

Section: Resultsmentioning

confidence: 99%

“… 10 Mechanical force generated by shaking culture-induced chondrogenic differentiation via TGF-β and Wnt signaling in 3D iPSC constructs. 19 Tensile force upregulated scleraxis, inhibiting ossification in PDLCs through TGF-β-ephrin A2 signaling. 39 In human gingival fibroblasts, compressive force increased cell proliferation, ECM synthesis and TGF-β expression, and a TGF-β inhibitor attenuated these effects.…”

Section: Discussionmentioning

confidence: 99%

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Intermittent compressive force induces cell cycling and reduces apoptosis in embryoid bodies of mouse induced pluripotent stem cells

et al. 2022

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In vitro manipulation of induced pluripotent stem cells (iPSCs) by environmental factors is of great interest for three-dimensional (3D) tissue/organ induction. The effects of mechanical force depend on many factors, including force and cell type. However, information on such effects in iPSCs is lacking. The aim of this study was to identify a molecular mechanism in iPSCs responding to intermittent compressive force (ICF) by analyzing the global gene expression profile. Embryoid bodies of mouse iPSCs, attached on a tissue culture plate in 3D form, were subjected to ICF in serum-free culture medium for 24 h. Gene ontology analyses for RNA sequencing data demonstrated that genes differentially regulated by ICF were mainly associated with metabolic processes, membrane and protein binding. Topology-based analysis demonstrated that ICF induced genes in cell cycle categories and downregulated genes associated with metabolic processes. The Kyoto Encyclopedia of Genes and Genomes database revealed differentially regulated genes related to the p53 signaling pathway and cell cycle. qPCR analysis demonstrated significant upregulation of Ccnd1, Cdk6 and Ccng1. Flow cytometry showed that ICF induced cell cycle and proliferation, while reducing the number of apoptotic cells. ICF also upregulated transforming growth factor β1 (Tgfb1) at both mRNA and protein levels, and pretreatment with a TGF-β inhibitor (SB431542) prior to ICF abolished ICF-induced Ccnd1 and Cdk6 expression. Taken together, these findings show that TGF-β signaling in iPSCs enhances proliferation and decreases apoptosis in response to ICF, that could give rise to an efficient protocol to manipulate iPSCs for organoid fabrication.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Intermittent compressive force induces cell cycling and reduces apoptosis in embryoid bodies of mouse induced pluripotent stem cells

et al. 2022

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“…Compressive stress also induced the chondrogenic commitment of iPSCs and ESCs via force‐activated TGF‐β overexpression and downstream Wnt signaling. [ 74 ]…”

Section: Biophysical Cues Of Biomaterials For Regulating Stem Cell Behaviormentioning

confidence: 99%

Manipulation of Stem Cells Fates: The Master and Multifaceted Roles of Biophysical Cues of Biomaterials

Wan

Liu

2021

Adv Funct Materials

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Owing to their self‐renewal and differentiation ability, stem cells are conducive for repairing injured tissues, making them a promising source of seed cells for tissue engineering. The extracellular microenvironment (ECM) is under dynamic mechanical control, which is closely related to stem cell behaviors. During the design and fabrication of biomaterials for regenerative medicine, the physiochemical properties of the natural ECM should be closely mimicked, which can reinforce stem cell lineage choice and tissue engineering. By reproducing the biophysical stimulations that stem cells may experience in vivo, many studies have highlighted the key role of biophysical cues in regulation of cell fate. Optimization of biophysical factors leads to desirable stem cell functions, which can maximize the effectiveness of regenerative treatment. In this review, the main biophysical cues of biomaterials, including stiffness, topography, mechanical force, and external physical fields are summarized, and their individual and synergistic influence on stem cell behavior is discussed. Subsequently, the current progress in tissue regeneration using biomaterials is presented, which directs the design and fabrication of functional biomaterial. The mechanisms via which biophysical cues activate cellular responses are also analyzed. Finally, the challenges in basic research as well as for clinical translation in this field are discussed.

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“…Thus, in some cases shear stress is positive: see-saw shaking of induced pluripotent stem cell (iPSC) constructs for 17 days promoted cell aggregation, and induced significantly higher expression of chondrogenic-related marker genes than observed in static cultures [15]. A platform rocking at 7 ° with a 3 second cycle results in an average shear stress of about 0.01 Pascal [16].…”

Section: Shear Stressmentioning

confidence: 99%

A Purpose-Built System for Culturing Cells as In Vivo Mimetic 3D Structures

Wrzesinski¹,

Alnøe²,

Jochumsen³

et al. 2021

Biomechanics and Functional Tissue Engineering

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Culturing cells in 3D is often considered to be significantly more difficult than culturing them in 2D. In practice, this is not the case: the situation is that equipment needed for 3D cell culture has not been optimised as much as equipment for 2D. Here we present a few key features which must be considered when designing 3D cell culture equipment. These include diffusion gradients, shear stress and time. Diffusion gradients are unavoidably introduced when cells are cultured as clusters. Perhaps the most important consequence of this is that the resulting hypoxia is a major driving force in the metabolic reprogramming. Most cells in tissues do not experience liquid shear stress and it should therefore be minimised. Time is the factor that is most often overlooked. Cells, irrespective of their origin, are damaged when cultures are initiated: they need time to recover. All of these features can be readily combined into a clinostat incubator and bioreactor. Surprisingly, growing cells in a clinostat system do not require specialised media, scaffolds, ECM substitutes or growth factors. This considerably facilitates the transition to 3D. Most importantly, cells growing this way mirror cells growing in vivo and are thus valuable for biomedical research.

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Shaking culture enhances chondrogenic differentiation of mouse induced pluripotent stem cell constructs

Cited by 21 publications

References 67 publications

Intermittent compressive force induces cell cycling and reduces apoptosis in embryoid bodies of mouse induced pluripotent stem cells

Intermittent compressive force induces cell cycling and reduces apoptosis in embryoid bodies of mouse induced pluripotent stem cells

Manipulation of Stem Cells Fates: The Master and Multifaceted Roles of Biophysical Cues of Biomaterials

A Purpose-Built System for Culturing Cells as In Vivo Mimetic 3D Structures

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