Spontaneous hepatocyte migration towards an endothelial cell tube network

Baimakhanov, Zhassulan; Sakai, Yusuke; Yamanouchi, Kaoru; Hidaka, Masaaki; Soyama, Akihiko; Takatsuki, Mitsuhisa; Eguchi, Susumu

doi:10.1002/term.2577

Cited by 5 publications

(7 citation statements)

References 12 publications

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“…It has been reported in several studies that the co‐culture model of endothelial cells and the other cells associated with diseases can simulate the phenotypic and morphological characteristics in vivo. Baimakhanov et al 31 observed that primary hepatocytes migrated spontaneously to the HUVEC tube network, and they hypothesized that this co‐culture model duplicated part of their normal physiology. In this study, HUVEC was used to simulate uterine spiral artery endothelium.…”

Section: Discussionmentioning

confidence: 99%

Effect of lncRNA‐ATB/miR‐651‐3p/Yin Yang 1 pathway on trophoblast‐endothelial cell interaction networks

Yin

Zhang

et al. 2021

J Cellular Molecular Medi

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Our previous studies have confirmed that lncRNA‐ATB may be involved in the pathogenesis of preeclampsia, however, it is uncertain whether lncRNA‐ATB influence the interaction between trophoblast and endothelial cells, which is crucial to the uterine spiral artery remodelling. Scratch wound healing and transwell invasion assay were conducted to test the migration and invasion of trophoblast cells. Co‐culture model was used to simulate the physiological environment in vivo. The expression levels of lncRNA‐ATB were analyzed in placenta tissues from healthy pregnant women and preeclampsia patients. Subsequently, the binding site of lncRNA‐ATB and miR‐651‐3p was verified using dual‐luciferase reporter assay, and the rescue experiment was used to study the effects of these two on the biological function. The direct effects of miR‐651‐3p and Yin Yang 1 (YY1) were verified using similar methods. LncRNA‐ATB was found to be down‐regulated in the placenta of preeclampsia patients. LncRNA‐ATB knockdown decreased trophoblast migration, invasion and colocalisation with human umbilical vein endothelial cells. MiR‐651‐3p was a direct target of lncRNA‐ATB and they had opposite effects. Moreover, the expression of lncRNA‐ATB and miR‐651‐3p in placental tissues was negatively correlated. MiR‐651‐3p has been confirmed to directly target the 3′ untranslated region of YY1. The inhibitory effects of YY1 low expression on biological function was rescued by miR‐651‐3p depletion. Western blot analysis showed that lncRNA‐ATB could regulate YY1 expression by sponging miR‐651‐3p. LncRNA‐ATB functioned as a competitive endogenous RNA of miR‐651‐3p to regulate YY1 on progress of spiral artery remodelling.

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Section: Discussionmentioning

confidence: 99%

Effect of lncRNA‐ATB/miR‐651‐3p/Yin Yang 1 pathway on trophoblast‐endothelial cell interaction networks

Yin

Zhang

et al. 2021

J Cellular Molecular Medi

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“…Previous studies have shown that hepatocytes contributed to the process of liver fibrosis through changing migratory behavior and biological functions. Hepatocytes can undergo epithelial-mesenchymal phenotypic transition, which increase the migratory ability and changes in the mechanical properties of hepatocytes can regulate cell migration and mechanical signal transduction [ 14 , 15 , 16 ]. However, at present, whether matrix stiffness, a factor that significantly changes in the process of liver fibrosis, can regulate hepatocytes participate in the process of liver fibrosis is unknown.…”

Section: Introductionmentioning

confidence: 99%

The Effect of Matrix Stiffness on Human Hepatocyte Migration and Function—An In Vitro Research

et al. 2020

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The extracellular matrix (ECM) regulates cellular function through the dynamic biomechanical and biochemical interplay between the resident cells and their microenvironment. Pathologically stiff ECM promotes phenotype changes in hepatocytes during liver fibrosis. To investigate the effect of ECM stiffness on hepatocyte migration and function, we designed an easy fabricated polyvinyl alcohol (PVA) hydrogel in which stiffness can be controlled by changing the concentration of glutaraldehyde. Three stiffnesses of hydrogels corresponding to the health of liver tissue, early stage, and end stage of fibrosis were selected. These were 4.8 kPa (soft), 21 kPa (moderate), and 45 kPa (stiff). For hepatocytes attachment, the hydrogel was coated with fibronectin. To evaluate the optimal concentration of fibronectin, hydrogel was coated with 0.1 mg/mL, 0.01 mg/mL, 0.005 mg/mL, or 0.003 mg/mL fibronectin, and the migratory behavior of single hepatocyte cultured on different concentrations of fibronectin was analyzed. To further explore the effect of substrate stiffness on hepatocyte migration, we used a stiffness controllable commercial 3D collagen gel, which has similar substrate stiffness to that of PVA hydrogel. Our result confirmed the PVA hydrogel biocompatibility with high hepatocytes survival. Fibronectin (0.01 mg/mL) promoted optimal migratory behavior for single hepatocytes. However, for confluent hepatocytes, a stiff substrate promoted hepatocellular migration compared with the soft and moderate groups via enhancing the formation of actin- and tubulin-rich structures. The gene expression analysis and protein expression analysis showed that the stiff substrate altered the phenotype of hepatocytes and induced apoptosis. Hepatocytes in stiff 3D hydrogel showed a higher proportion of cell death and expression of filopodia.

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“…Many studies utilize DMEM for the culture of thyroid cell lines [17]. While HDM was originally designed for the culture of hepatocytes, it has also been reported to be suitable for the culture of epithelial cells and certain kinds of gland cells, such as hepatic epithelial cells [15], salivary epithelial cells [18] and primary human lacrimal gland cells [19]. Similarly, thyroid cells, which are follicle epithelial cells isolated from the thyroid gland, may also be well-cultured and easily proliferate in epithelial cell culture medium.…”

Section: Discussionmentioning

confidence: 99%

“…Because no chemical treatments are needed in order to harvest the cell sheets with extracellular matrix, the native cellular function is maintained [4,6,15]. Furthermore, cell sheets have been shown to be able to be handled easily.…”

Section: Discussionmentioning

confidence: 99%

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Fabrication of Functional Cell Sheets with Human Thyrocytes from Non-Tumorous Thyroid Tissue

Huang

Yamanouchi

Sakai

et al. 2019

Tissue Eng Regen Med

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BACKGROUND: Engineered cell sheet transplantation has been considered an alternative physiological therapy for endocrine disorders. In this study, we attempted to fabricate functional human thyroid cell sheets using the engineering technology by culturing primary thyrocytes in free-feeder monolayers and assessed their proliferation and function in two different media. METHODS: The non-tumorous tissues (approximately 2 g) were dissected during surgery. Primary human thyroid cells were isolated by mechanical dispersion and treatment with isolation solution. The cells were cultured on tissue culture dishes or temperature-responsive culture dishes to induce the formation of detached cell sheets. RESULTS: Primary thyroid cells isolated from nine patients were positive for thyroid transcription factor 1, thyroglobulin (TG) and cytokeratin 7. Cell sheets with follicles were fabricated by cells incubated in both Dulbecco's Modified Eagle Medium (DMEM) and hepatocyte-defined medium (HDM) culture medium. The diameter and thickness of sheets fabricated in HDM were larger and thicker than those fabricated from DMEM. Furthermore, the cells incubated in HDM secreted higher levels of fT3 and fT4 than those incubated in DMEM. The thyroid peroxidase and TG mRNA of cells maintained in HDM were higher than those in cells maintained in DMEM. CONCLUSION: HDM appears suitable as a culture medium for maintaining primary thyrocytes and fabricating functional cell sheets. These in vitro findings may contribute to the development of appropriate culture conditions for human thyrocytes as well as engineered functional cell sheets.

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Spontaneous hepatocyte migration towards an endothelial cell tube network

Cited by 5 publications

References 12 publications

Effect of lncRNA‐ATB/miR‐651‐3p/Yin Yang 1 pathway on trophoblast‐endothelial cell interaction networks

Effect of lncRNA‐ATB/miR‐651‐3p/Yin Yang 1 pathway on trophoblast‐endothelial cell interaction networks

The Effect of Matrix Stiffness on Human Hepatocyte Migration and Function—An In Vitro Research

Fabrication of Functional Cell Sheets with Human Thyrocytes from Non-Tumorous Thyroid Tissue

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