Kyushu University Academic Staff Educational and Research Activities Database
List of Presentations
THASANEEYA KUBOKI Last modified date:2020.07.02

Assistant Professor / Laboratory of Biomedical and Biophysical Chemistry / Department of Applied Molecular Chemistry / Institute for Materials Chemistry and Engineering


Presentations
1. Kuboki Thasaneeya, Kantawong Fahsai, Kidoaki Satoru, Effects of mechanical stimuli on redox homeostasis of mesenchymal stem cells, Office for the promotion of Gender equality, 2019.03.
2. Kuboki Thasaneeya, Mechanotransduction and redox regulation of stem cells, Selectbio, 2018.07.
3. Kuboki Thasaneeya, Kantawong Fahsai, Kidoaki Satoru, Mechanotransduction and redox regulation of stem cells., STEM18, 2018.03.
4. THASANEEYA KUBOKI, Fahsai Kantawong, Satoru Kidoaki, Mechanotransduction and redox regulation of stem cells., Biomaterials International 2017 , 2017.08.
5. THASANEEYA KUBOKI, Fahsai Kantawong, Satoru Kidoaki, Mechanotransduction and redox regulation of stem cells., ISB2017, 2017.07.
6. THASANEEYA KUBOKI, Fahsai Kantawong, Satoru Kidoaki, Mechanotransduction and redox signaling of stem cells., Mechanobiology of Disease , 2016.09.
7. THASANEEYA KUBOKI, Satoru Kidoaki, Live imaging of paxillin in durotactic migrating cells on the micro-elastically patterned hydrogels., KJF-ICOM , 2016.09.
8. THASANEEYA KUBOKI, Satoru Kidoaki, Surface elasticity tunable gelatinous gel for manipulation of stem cell fate determination and directional cell migration., PCT, 2016.06.
9. KUBOKI THASANEEYA, Japan Society of Mechanical Engineers 28; Bioengineering lecture , 2016.01, The mechanism of durotaxis was characterized using gelatinous square domain patterned gel on the soft base (300/40 kPa). The Fluorescence Recovery After Photobleaching analysis of 3T3 fiborblasts expressing venus-paxillin showed the stiffness, time and position dependent mobility of paxillin in the durotactic migrating cells. Time-lapse observation provided information concerning rate of assembly/disassembly and turnover of paxillin at the surface elasticity boundaries. .
10. Kantawong F, Kuboki T, Kidoaki S, REDOX GENE EXPRESSION OF ADIPOSE-DERIVED STEM CELLS IN RESPONSE TO SOFT HYDROGEL, The 8th Asian-Pacific Conference on Biomechanics (AP-Biomech 2015), 2015.09, Introduction
Adipose-derived stem cells (ADSCs) are multipotent stem cells within the adipose tissue, which are considered as a promising source of stem cell population. ADSCs offer unique opportunities as novel cell-based therapeutics and as traditional pharmaceutical discovery tools, which could have a significant therapeutic impact in the future.
When cultured on very soft surface ADSCs showed morphological change to a neuron-like shape, and presented neuronal lineage bias. To gain the basic insights on gene expression relating to the neuronal lineage bias in ADSCs, we examined the correlation between the gene expression levels of neuronal markers and redox proteins that have recently been considered to have close relation with lineage specification in stem cells.
Methods
The surface elasticity tunable hydrogel was fabricated using photocurable styrenated gelatin. The elasticity of the gelatinous gel was measured using atomic force microscope and the 4 kPa hydrogels were used for ADSCs cultured for 4 days to 2 weeks. The time course expression of neuronal genes (TUBB3 and NSE) and redox genes (TRX1, SOD1, SOD2, PRX2, GSTT1, and GSTP1) were monitored using real-time PCR.
Result and discussion
It was found that the TUBB3 gene had significantly up-regulated, compared with the control tissue culture polystyrene, indicating that the neuronal gene expression of ADSCs could be achieved on soft hydrogel without the addition of any supplement. The expressions of the TRX1 and SOD1 genes were also observed to have significantly up-regulated on the soft hydrogels. The results demonstrate that the up-regulation of the neural marker of TUBB3 in the ADSCs correlates well with the up-regulation of the redox genes of TRX1 and SOD1 when cultured on appropriate soft hydrogel substrates. The changes in expression of these redox genes might have roles to play in the expression of neuronal markers, or they could be a response to the oxidative stress caused by the hydrogels, regarding which further studies are required.
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11. THASANEEYA KUBOKI, Time-dependent migratory behaviors in the long-term studies of fibroblast durotaxis on a hydrogel substrate fabricated with a soft band , Mechanobiology, 2014.05, [URL], Durotaxis, biased cell movement up a stiffness gradient on culture substrates, is one of the useful taxis behaviors for manipulating cell migration on engineered biomaterial surfaces. In this study, long-term durotaxis was investigated on gelatinous substrates containing a soft band of 20 µm, 50 µm, and 150 µm in width fabricated using photolithographic elasticity patterning; sharp elasticity boundaries with a gradient strength of 300 kPa/50 µm were achieved. Time-dependent migratory behaviors of 3T3 fibroblast cells were observed during a time period of three days. During the first day, most of the cells were strongly repelled by the soft band independent of band-width, exhibiting the typical durotaxis behavior. However, the repellency by the soft band diminished and more cells crossed the soft band or exhibited other mixed migratory behaviors during the course of the observation. It was found that durotaxis strength is weakened on the substrate with the narrowest soft band and that adherent affinity-induced entrapment becomes apparent on the widest soft band with time. Factors, such as changes in surface topography, elasticity, and/or chemistry, likely contributing to the apparent diminishing durotaxis during the extended culture were examined. Immunofluorescence analysis indicated preferential collagen deposition onto the soft band, which is derived from secretion by fibroblast cells, resulting in the increasing contribution of haptotaxis toward the soft band over time. The deposited collagen did not affect surface topography or surface elasticity, but did change surface chemistry, especially on the soft band. The observed time-dependent durotaxis behaviors are the result of the mixed mechanical and chemical cues. In the studies and applications of cell migratory behavior under a controlled stimulus, it is important to thoroughly examine other (hidden) compounding stimuli in order to be able to accurately interpret data and to design suitable biomaterials to manipulate cell migration. .
12. Kuboki T, Chen W, Kidoaki S, Controlling mechano-repellent cell migration induced around a micro-scale soft stripe on hydrogel matrix. , International Nanomedicine Conference, (Sydney, Australia), 2013.07, Cell migration is a fundamental aspect of many physiological and pathological processes such as embryonic development, tissue morphogenesis, wound healing and cancer metastasis. Various factors in cellular microenvironment participate in the regulation of cell migration including soluble factors and mechanical stimuli from extracellular matrix. Fabrication of mechanically patterned substrates is essential in the understanding of how cell migration is affected by mechanical cues.
Mechanotaxis or durotaxis describes the phenomenon that cells preferentially migrate toward stiffer domains on a substrate where a mechanical gradient is present 1. Our study focused on surface elasticity-induced directional cell migration. We demonstrated the feasibility in controlling directional cell migration, i.e. turning or repelling, based on mechanotaxis using patterned gels containing a single soft stripe (Fig. 1).
The photocurable styrenated gelatin 2, 3, was used for the fabrication of patterned gels containing a narrow soft stripe (20, 50 or 150 µm) against a stiffer background, which was prepared using a newly developed Liquid Crystal Display (LCD) projector photolithographic patterning method 4. The surface elasticity of the hard domains (400 kPa) and the soft stripes (80 kPa) were designed according to the condition required to induce mechanotaxis 3. Repellency of 3T3 fibroblasts migrating from the hard domain by the soft stripe was observed in a time-dependent manner. The narrowest 20-µm wide stripes induced the strongest cell repellency, and similar trends were observed in conditions of stripe with three different widths.
During the first day of time-lapse observations, majority of the cells were repelled upon approaching the soft regions. After the second and third days, fewer cells were repelled and more cells managed to cross the soft stripes. No noticeable changes in surface topography and gel elasticity after prolong cell cultured were observed.
It was speculated that the time-dependent cell migratory behaviours were attributed to the secreted collagen deposited over time at the elasticity boundaries, as the fibroblasts are known to produce collagen. Immunofluorescense staining with anti-collagen I indicated that collagen deposition is a likely contributor to the changes in the cell repellency by the soft stripes over the time.
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