• Title/Summary/Keyword: hMSC

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Effects of $CoCl_2$ on Osteogenic Differentiation of Human Mesenchymal Stem Cells

  • Moon, Yeon-Hee;Son, Jung-Wan;Moon, Jung-Sun;Kang, Jee-Hae;Kim, Sun-Hun;Kim, Min-Seok
    • International Journal of Oral Biology
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    • v.38 no.3
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    • pp.111-119
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    • 2013
  • Objective. To investigate the effects of the hypoxia inducible factor-1 (HIF-1) activation-mimicking agent cobalt chloride ($CoCl_2$) on the osteogenic differentiation of human mesenchymal stem cells (hMSCs) and elucidate the underlying molecular mechanisms. Study design. The dose and exposure periods for $CoCl_2$ in hMSCs were optimized by cell viability assays. After confirmation of $CoCl_2$-induced HIF-$1{\alpha}$ and vascular endothelial growth factor expression in these cells by RT-PCR, the effects of temporary preconditioning with $CoCl_2$ on hMSC osteogenic differentiation were evaluated by RT-PCR analysis of osteogenic gene expression, an alkaline phosphatase (ALP) activity assay and by alizarin red S staining. Results. Variable $CoCl_2$ dosages (up to $500{\mu}M$) and exposure times (up to 7 days) on hMSC had little effect on hMSC survival. After $CoCl_2$ treatment of hMSCs at $100{\mu}M$ for 24 or 48 hours, followed by culture in osteogenic differentiating media, several osteogenic markers such as Runx-2, osteocalcin and osteopontin, bone sialoprotein mRNA expression level were found to be up-regulated. Moreover, ALP activity was increased in these treated cells in which an accelerated osteogenic capacity was also verified by alizarin red S staining. Conclusions. The osteogenic differentiation potential of hMSCs could be preserved and even enhanced by $CoCl_2$ treatment.

Real-time Control of Biological Animal Wastewater Treatment Process and Stability of Control Parameters (생물학적 축산폐수 처리공정의 자동제어 방법 및 제어 인자의 안정성)

  • Kim, W.Y.;Jung, J.H.;Ra, C.S.
    • Journal of Animal Science and Technology
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    • v.46 no.2
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    • pp.251-260
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    • 2004
  • The feasibility and stability of ORP, pH(mV) and DO as a real-time control parameter for SBR process were evaluated in this study. During operation, NBP(nitrogen break point) and NKP(nitrate knee point), which reveal the biological and chemical changes of pollutants, were clearly observed on ORP and pH(mV)-time profiles, and those control points were easily detected by tracking the moving slope changes(MSC). However, when balance of aeration rate to loading rate, or to OUR(oxygen uptake rate), was not optimally maintained, either false NBP was occurred on ORP and DO curves before the appearance of real NBP or specific NBP feature was disappeared on ORP curve. Under that condition, however, very distinct NBP was found on pH(mV)-time profile, and stable detection of that point was feasible by tracking MSC. These results might mean that pH(mV) is superior real-time control parameter for aerobic process than ORP and DO. Meanwhile, as a real-time control parameter for anoxic process, ORP was very stable and more useful parameter than others. Based on these results, a stable real-time control of process can be achieved by using the ORP and pH(mv) parameters in combination rather than using separately. A complete removal of pollutants could be always ensured with this real-time control technology, despite the variations of wastewater and operation condition, as well as an optimization of treatment time and capacity could be feasible.

Two-Cell Spheroid Angiogenesis Assay System Using Both Endothelial Colony Forming Cells and Mesenchymal Stem Cells

  • Shah, Sajita;Kang, Kyu-Tae
    • Biomolecules & Therapeutics
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    • v.26 no.5
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    • pp.474-480
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    • 2018
  • Most angiogenesis assays are performed using endothelial cells. However, blood vessels are composed of two cell types: endothelial cells and pericytes. Thus, co-culture of two vascular cells should be employed to evaluate angiogenic properties. Here, we developed an in vitro 3-dimensional angiogenesis assay system using spheroids formed by two human vascular precursors: endothelial colony forming cells (ECFCs) and mesenchymal stem cells (MSCs). ECFCs, MSCs, or ECFCs+MSCs were cultured to form spheroids. Sprout formation from each spheroid was observed for 24 h by real-time cell recorder. Sprout number and length were higher in ECFC+MSC spheroids than ECFC-only spheroids. No sprouts were observed in MSC-only spheroids. Sprout formation by ECFC spheroids was increased by treatment with vascular endothelial growth factor (VEGF) or combination of VEGF and fibroblast growth factor-2 (FGF-2). Interestingly, there was no further increase in sprout formation by ECFC+MSC spheroids in response to VEGF or VEGF+FGF-2, suggesting that MSCs stimulate sprout formation by ECFCs. Immuno-fluorescent labeling technique revealed that MSCs surrounded ECFC-mediated sprout structures. We tested vatalanib, VEGF inhibitor, using ECFC and ECFC+MSC spheroids. Vatalanib significantly inhibited sprout formation in both spheroids. Of note, the $IC_{50}$ of vatalanib in ECFC+MSC spheroids at 24 h was $4.0{\pm}0.40{\mu}M$, which are more correlated with the data of previous animal studies when compared with ECFC spheroids ($0.2{\pm}0.03{\mu}M$). These results suggest that ECFC+MSC spheroids generate physiologically relevant sprout structures composed of two types of vascular cells, and will be an effective pre-clinical in vitro assay model to evaluate pro- or anti-angiogenic property.

Change in Concentrations of Human Norovirus and Male-Specific Coliphage under Various Temperatures, Salinities, and pH Levels in Seawater (해수 중의 수온, 염분 및 pH에 따른 노로바이러스 및 Male-Specific Coliphage 농도변화)

  • Kim, Poong Ho;Park, Yong Soo;Park, Kunbawui;Kwon, Ji Young;Yu, Hong Sik;Lee, Hee Jung;Kim, Ji Hoe;Lee, Tae Seek
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.4
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    • pp.454-459
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    • 2016
  • Pre- or post-harvest processing is required to mitigate the risk of norovirus infection mediated by shellfish or seafood. We investigated the environmental resistance of human norovirus (HuNoV) under various conditions of temperature, salinity, and pH in seawater. Male-specific coliphage (MSC) was as the reference virus for all tests. At 4℃, HuNoV GII4 spiked into seawater was continually detected by RT-PCR for 35 days, regardless of salinity or pH level. It maintained nearly stable concentrations, meaning HuNoV can sustain a viral population in seawater long enough to be accumulated by shellfish and other filter feeders during winter. MSC was also stable at 4℃ although viral infectivity dropped sharply after 28 days. The effects of salinity and pH on MSC were indistinct. At 25℃ the detectable period of HuNoV GII4 by RT-PCR in seawater decreased to about one-third or half of the period at 4℃. High salinity (32 psu) and alkaline pH (8.5) were also unfavorable for sustaining HuNoV abundance at 25℃ in seawater. The resistance patterns of MSC to high temperature, high salinity, and alkaline pH were more dramatic and viral infectivity decreased over time, almost in direct proportion to experimental days. MSC was undetectable after 12 days under all salinities and pH levels at 25℃.

Analysis on the Measurement Results of the Focus Motor Position in MSC (Multi-Spectral Camera) on KOMPSAT - II

  • Heo, H.P.;Kong, J.P.;Kim, Y.S.;Park, J.E.;Chang, Y.J.;Lee, S.H.
    • Proceedings of the KSRS Conference
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    • v.1
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    • pp.372-375
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    • 2006
  • The MSC is a high resolution multi-spectral camera system which is mounted on the KOMPSAT-II satellite. The electro-optic camera system has a refocusing mechanism which can be used in-orbit by ground commands. By adjusting locations of some elements in optics, the system can be focused precisely. The focus mechanism in MSC is implemented with stepper motor and potentiometer. By reading the value of the potentiometer, rough position of the motor can be understood. The exact location of the motor can not be acquired because the information from the potentiometer can not be so accurate. However, before and after certain events of the satellite, like a satellite launch, the direction of the movement or order of the magnitude of the movement can be understood. In this paper, the trend analysis of the focus motor position during the ground test phase is introduced. This result can be used as basic information for the focus calibration after launch. By studying the long term trend, deviation from the best focal point can be understood. The positions of the focus motors after launch are also compared.

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$PKC{\eta}$ Regulates the $TGF{\beta}3$-induced Chondrogenic Differentiation of Human Mesenchymal Stem Cell

  • Ku, Bo Mi;Yune, Young Phil;Lee, Eun Shin;Hah, Young-Sool;Park, Jae Yong;Jeong, Joo Yeon;Lee, Dong Hoon;Cho, Gyeong Jae;Choi, Wan Sung;Kang, Sang Soo
    • Development and Reproduction
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    • v.17 no.4
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    • pp.299-309
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    • 2013
  • Transforming growth factor (TGF) family is well known to induce the chondrogenic differentiation of mesenchymal stem cells (MSC). However, the precise signal transduction pathways and underlying factors are not well known. Thus the present study aims to evaluate the possible role of C2 domain in the chondrogenic differentiation of human mesenchymal stem cells. To this end, 145 C2 domains in the adenovirus were individually transfected to hMSC, and morphological changes were examined. Among 145 C2 domains, C2 domain of protein kinase C eta ($PKC{\eta}$) was selected as a possible chondrogenic differentiation factor for hMSC. To confirm this possibility, we treated $TGF{\beta}3$, a well known chondrogenic differentiation factor of hMSC, and examined the increased-expression of glycosaminoglycan (GAG), collagen type II (COL II) as well as $PKC{\eta}$ using PT-PCR, immunocytochemistry and Western blot analysis. To further evaluation of C2 domain of $PKC{\eta}$, we examined morphological changes, expressions of GAG and COL II after transfection of $PKC{\eta}$-C2 domain in hMSC. Overexpression of $PKC{\eta}$-C2 domain induced morphological change and increased GAG and COL II expressions. The present results demonstrate that $PKC{\eta}$ involves in the TGF-${\beta}3$-induced chondrogenic differentiation of hMSC, and C2 domain of $PKC{\eta}$ has important role in this process.

Nanoengineered, cell-derived extracellular matrix influences ECM-related gene expression of mesenchymal stem cells

  • Ozguldez, Hatice O.;Cha, Junghwa;Hong, Yoonmi;Koh, Ilkyoo;Kim, Pilnam
    • Biomaterials Research
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    • v.22 no.4
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    • pp.337-345
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    • 2018
  • Background: Human mesenchymal stem cells (hMSCs) are, due to their pluripotency, useful sources of cells for stem cell therapy and tissue regeneration. The phenotypes of hMSCs are strongly influenced by their microenvironment, in particular the extracellular matrix (ECM), the composition and structure of which are important in regulating stem cell fate. In reciprocal manner, the properties of ECM are remodeled by the hMSCs, but the mechanism involved in ECM remodeling by hMSCs under topographical stimulus is unclear. In this study, we therefore examined the effect of nanotopography on the expression of ECM proteins by hMSCs by analyzing the quantity and structure of the ECM on a nanogrooved surface. Methods: To develop the nanoengineered, hMSC-derived ECM, we fabricated the nanogrooves on a coverglass using a UV-curable polyurethane acrylate (PUA). Then, hMSCs were cultivated on the nanogrooves, and the cells at the full confluency were decellularized. To analyze the effect of nanotopography on the hMSCs, the hMSCs were re-seeded on the nanoengineered, hMSC-derived ECM. Results: hMSCs cultured within the nano-engineered hMSC-derived ECM sheet showed a different pattern of expression of ECM proteins from those cultured on ECM-free, nanogrooved surface. Moreover, hMSCs on the nano-engineered ECM sheet had a shorter vinculin length and were less well-aligned than those on the other surface. In addition, the expression pattern of ECM-related genes by hMSCs on the nanoengineered ECM sheet was altered. Interestingly, the expression of genes for osteogenesis-related ECM proteins was downregulated, while that of genes for chondrogenesis-related ECM proteins was upregulated, on the nanoengineered ECM sheet. Conclusions: The nanoengineered ECM influenced the phenotypic features of hMSCs, and that hMSCs can remodel their ECM microenvironment in the presence of a nanostructured ECM to guide differentiation into a specific lineage.

MSC(Multi-Spectral Camera) 열제어 시스템 소개

  • Kong, Jong-Pil;Heo, Haeng-Pal;Kim, Young-Sun;Park, Jong-Euk;Jang, Young-Jun
    • Aerospace Engineering and Technology
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    • v.4 no.2
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    • pp.107-116
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    • 2005
  • As a unique payload of Komsat-2, MSC, comprising EOS(Electro-Optical Sub-system), PMU(Payload Management Unit) and PDTS(Payload Data Transmission Sub-system), is supposed to take pictures of one panchromatic and 4 multi-spectral image between wavelength 450mm~900mm, and is being under final Satellite I&T. It will perform the earth remote sensing with applications such as acquisition of high resolution images, surveillance of large scale disasters and its countermeasure, survey of natural resources, etc.. Under the hostile influence of the extreme space environmental conditions due to deep space and direct solar flux, the thermal design is especially of major importance in designing a payload. There are tight temperature range restrictions for electro-optical elements while on the other hand there are low power consumption requirements due to the limited energy source on the spacecraft. This paper describes details of thermal control system for MSC.

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Construction of Global Finite State Machine from Message Sequence Charts for Testing Task Interactions (태스크 상호작용 테스팅을 위한 MSC 명세로부터의 전체 유한 상태 기계 생성)

  • Lee, Nam-Hee;Kim, Tai-Hyo;Cha, Sung-Deok;Shin, Seog-Jong;Hong, H-In-Pyo;Park, Ki-Wung
    • Journal of KIISE:Software and Applications
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    • v.28 no.9
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    • pp.634-648
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    • 2001
  • Message Sequence Charts(MSC) has been used to describe the interactions of numerous concurrent tasks in telecommunication software. After the MSC specification is verified in requirement analysis phase, it can be used not only to synthesize state-based design models, but also to generate test sequences. Until now, the verification is accomplished by generating global state transition graph using the location information only. In this paper, we extend the condition statement of MSC to describe the activation condition of scenarios and the change of state variables, and propose an approach to construct global finite state machine (GFSM) using this information. The GFSM only includes feasible states and transitions of the system. We can generate the test sequences using the existing FSM-based test sequence generation technology.

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Effects of Human Mesenchymal Stem Cell Transplantation Combined with Polymer on Functional Recovery Following Spinal Cord Hemisection in Rats

  • Choi, Ji Soo;Leem, Joong Woo;Lee, Kyung Hee;Kim, Sung-Soo;SuhKim, Haeyoung;Jung, Se Jung;Kim, Un Jeng;Lee, Bae Hwan
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.6
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    • pp.405-411
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    • 2012
  • The spontaneous axon regeneration of damaged neurons is limited after spinal cord injury (SCI). Recently, mesenchymal stem cell (MSC) transplantation was proposed as a potential approach for enhancing nerve regeneration that avoids the ethical issues associated with embryonic stem cell transplantation. As SCI is a complex pathological entity, the treatment of SCI requires a multipronged approach. The purpose of the present study was to investigate the functional recovery and therapeutic potential of human MSCs (hMSCs) and polymer in a spinal cord hemisection injury model. Rats were subjected to hemisection injuries and then divided into three groups. Two groups of rats underwent partial thoracic hemisection injury followed by implantation of either polymer only or polymer with hMSCs. Another hemisection-only group was used as a control. Behavioral, electrophysiological and immunohistochemical studies were performed on all rats. The functional recovery was significantly improved in the polymer with hMSC-transplanted group as compared with control at five weeks after transplantation. The results of electrophysiologic study demonstrated that the latency of somatosensory-evoked potentials (SSEPs) in the polymer with hMSC-transplanted group was significantly shorter than in the hemisection-only control group. In the results of immunohistochemical study, ${\beta}$-gal-positive cells were observed in the injured and adjacent sites after hMSC transplantation. Surviving hMSCs differentiated into various cell types such as neurons, astrocytes and oligodendrocytes. These data suggest that hMSC transplantation with polymer may play an important role in functional recovery and axonal regeneration after SCI, and may be a potential therapeutic strategy for SCI.