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표면-줄기세포 증식 관계 확인을 위한 마이크로 구조표면을 포함한 멀티웰 플랫폼 제작
안성아(Seonga An),최동휘(Dongwhi Choi),임지원(Jiwon Lim),김동성(Dong Sung Kim) 대한기계학회 2015 대한기계학회 춘추학술대회 Vol.2015 No.11
Micro-scale topography fabrication have attracted much attention in biomedical engineering because of their potential for stem cell proliferation enhancement effect. In recent years, adult stem cells (ASCs) are proved as versatile troubleshooter in biomedical and pharmaceutical field. However, ASCs take up only a small proportion of somatic cells in human body and the study of enhancing the proliferation rate of ASCs became essential in modern society. Researchers suggested the use of topographic stimuli as solution for this problem. Numerous researches were executed to clarify the relationship of topography and ASCs proliferation but there are still difficulties in defining tendency to clearly deduce an optimal topography for enhancing proliferation rate due to an intervention of other factors. To overcome this weakness we propose a multi-well cell culture platform that allows simple way of screening effects of various kinds of topography on ASCs. In this study we fabricated multi-well cell culture platform containing micro-scale topography and carried out experiments of human Adipose derived Stem Cells (hASCs) proliferation. Through this experiment we verify the multi-well cell culture platform’s functionality then we expect that this multi-well cell culture platform system would contribute to finding optimal topography for enhancing ASCs proliferation rate.
Autophagy alteration prevents primary cilium disassembly in RPE1 cells
Maharjan, Yunash,Lee, Joon No,Kwak, SeongAe,Lim, Hyewon,Dutta, Raghbendra Kumar,Liu, Zhi-qiang,So, Hong-Seob,Park, Raekil Elsevier 2018 Biochemical and biophysical research communication Vol.500 No.2
<P><B>Abstract</B></P> <P>Primary cilium is a microtubule structure that emanates from the surface of most human cells. Primary cilia assemble during the resting stage (G<SUB>0</SUB> phase) and disassemble with cell cycle progression. Defects associated with the control of the assembly or disassembly of the primary cilium have been implicated in various human diseases, including ciliopathy and cancer. Although studies have suggested the interplay between activation of autophagy and ciliogenesis, any direct mechanism between autophagy abatement and disassembly of primary cilium remains elusive. In this study, we found that the gradual abatement in autophagy during serum-restimulation was a dynamic process and significantly correlated with the disassembly of primary cilium in human retinal pigmented epithelial (RPE1) cells. Although autophagy activity was gradually decreased during serum-restimulation, the alteration in autophagy under the same condition prevented the disassembly of the primary cilium. Autophagy inhibitors such as chloroquine, U18666A and 3-methyladenine (3-MA) retained both the number of ciliated cells and cilium length. In contrast, rapamycin treatment during serum-restimulation maintained the number of ciliated cells with shortened cilia. Taken together, alteration in autophagy during serum-restimulation prevent the disassembly of the primary cilium, and autophagy modulators may serve as useful compounds for studying mechanistic details related to the disassembly of the primary cilium and ciliopathy.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Autophagy abatement and cilia disassembly is correlated during serum-restimulation. </LI> <LI> Both activation and inhibition of autophagy flux during serum-restimulation prevents disassembly of primary cilium. </LI> <LI> Inhibition of autophagy maintains cilium length during serum-restimulation. </LI> <LI> Rapamycin-induced autophagy results in shortened cilium length during serum-restimulation. </LI> </UL> </P>
Liu, Zhi-qiang,Lee, Joon No,Son, Myeongjoo,Lim, Jae-Young,Dutta, Raghbendra Kumar,Maharjan, Yunash,Kwak, SeongAe,Oh, Goo Taeg,Byun, Kyunghee,Choe, Seong-Kyu,Park, Raekil Landes Bioscience 2018 AUTOPHAGY Vol.14 No.6
<P>The primary cilia are evolutionarily conserved microtubule-based cellular organelles that perceive metabolic status and thus link the sensory system to cellular signaling pathways. Therefore, ciliogenesis is thought to be tightly linked to autophagy, which is also regulated by nutrient-sensing transcription factors, such as PPARA (peroxisome proliferator activated receptor alpha) and NR1H4/FXR (nuclear receptor subfamily 1, group H, member 4). However, the relationship between these factors and ciliogenesis has not been clearly demonstrated. Here, we present direct evidence for the involvement of macroautophagic/autophagic regulators in controlling ciliogenesis. We showed that activation of PPARA facilitated ciliogenesis independently of cellular nutritional states. Importantly, PPARA-induced ciliogenesis was mediated by controlling autophagy, since either pharmacological or genetic inactivation of autophagy significantly repressed ciliogenesis. Moreover, we showed that pharmacological activator of autophagy, rapamycin, recovered repressed ciliogenesis in ppara(-/-) cells. Conversely, activation of NR1H4 repressed cilia formation, while knockdown of NR1H4 enhanced ciliogenesis by inducing autophagy. The reciprocal activities of PPARA and NR1H4 in regulating ciliogenesis were highlighted in a condition where de-repressed ciliogenesis by NR1H4 knockdown was further enhanced by PPARA activation. The in vivo roles of PPARA and NR1H4 in regulating ciliogenesis were examined in greater detail in ppara(-/-) mice. In response to starvation, ciliogenesis was facilitated in wild-type mice via enhanced autophagy in kidney, while ppara(-/-) mice displayed impaired autophagy and kidney damage resembling ciliopathy. Furthermore, an NR1H4 agonist exacerbated kidney damage associated with starvation in ppara(-/-) mice. These findings indicate a previously unknown role for PPARA and NR1H4 in regulating the autophagy-ciliogenesis axis in vivo.</P>