• Title/Summary/Keyword: cell recovery

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In situ Recovery of hGM-CSF in Transgenic Rice Cell Suspension Cultures (형질전환 벼 현탁세포 배양에서 hGM-CSF의 in situ Recovery 연구)

  • Myoung, Hyun-Jong;Choi, Hong-Yeol;Nam, Hyung-Jin;Kim, Dong-Il
    • KSBB Journal
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    • v.30 no.3
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    • pp.103-108
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    • 2015
  • Production of foreign proteins by transgenic plant cell cultures has several advantages such as post-translational modification, low risk of product contamination and low-cost production and purification. However, target proteins are degraded by extracellular proteases existing in the media. A solution to this problem is the use of perfusion culture and ion exchange chromatography for the application of integrated bioprocess using in situ recovery. With this method, production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in this study. First, optimization of cell concentration during the induction phase for the production of hGM-CSF was examined. As cell concentration increased, the level of hGM-CSF was decreased due to the presence of extracellular proteases. Induction using sugarfree media produced 33% more hGM-CSF. The effects of pH on the binding of hGM-CSF to cationic and anionic exchange resins were also investigated. In terms of stability, optimal pH was found to be 5~7. In the case of using buffer exchange when CM-Sepharose was used as a cationic exchange resin, optimal pH for binding was 4.8 and adsorption yield was 77%. When DEAE-Sepharose was used as an anionic exchange resin, it was 5.5 (74%). Without buffer exchange, optimal pH was 4.6 (84%). From these results, an integrated bioprocess using in situ recovery with simultaneous production and separation of foreign protein in transgenic plant cell suspension cultures was found to be feasible.

Improvement of Functional Recovery by Cell Transplantation after Spinal Cord Injury (척수손상 후 세포이식에 의한 운동기능의 회복증진)

  • 이배환;이경희;성제경;황세진;김계성
    • Science of Emotion and Sensibility
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    • v.7 no.2
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    • pp.179-186
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    • 2004
  • Acute spinal cord injury can produce neurologic injury with many physical, psychological and social ramifications. It has been shown that two separate components combine to produce neurologic damage in acute spinal cord injury : the primary and secondary injuries. The primary mediators of spinal cord injury include the actual mechanical tissue disruption which is a passive process that occurs immediately following the trauma. A secondary injury cascade follows which appears mediated by cellular and molecular processes working through complex mechanisms. Both the primary and secondary injury cascades produce cell death both in neuronal and supporting cell tissues. Recovery from central nervous system(CNS) disorders is hindered by the limited ability of the vertebrate CNS to regenerate injured cells, replace damaged myelin sheath, and re-establish functional neuronal connections. Of many CNS disorders including multiple sclerosis, stroke, and other trauma, spinal cord injury is one of the important diseases because of the direct association with the functional loss of the body. Previous studies suggest that substantial recovery of function might be achieved through regeneration of lost neuronal cells and remyelination of intact axon in spinal cord injury which is occurred frequently. As a therapeutic approach in spinal cord injury, recently, cell transplantation provides a potential solution for the treatment of spinal cord injury. This review describes the characteristics of spinal cord injury and presents some evidence supporting functional recovery after cell transplantation following spinal cord injury.

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Reduction of Inflammation and Enhancement of Motility after Pancreatic Islet Derived Stem Cell Transplantation Following Spinal Cord Injury

  • Karaoz, Erdal;Tepekoy, Filiz;Yilmaz, Irem;Subasi, Cansu;Kabatas, Serdar
    • Journal of Korean Neurosurgical Society
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    • v.62 no.2
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    • pp.153-165
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    • 2019
  • Objective : Spinal cord injury (SCI) is a very serious health problem, usually caused by a trauma and accompanied by elevated levels of inflammation indicators. Stem cell-based therapy is promising some valuable strategies for its functional recovery. Nestin-positive progenitor and/or stem cells (SC) isolated from pancreatic islets (PI) show mesenchymal stem cell (MSC) characteristics. For this reason, we aimed to analyze the effects of rat pancreatic islet derived stem cell (rPI-SC) delivery on functional recovery, as well as the levels of inflammation factors following SCI. Methods : rPI-SCs were isolated, cultured and their MSC characteristics were determined through flow cytometry and immunofluorescence analysis. The experimental rat population was divided into three groups : 1) laminectomy & trauma, 2) laminectomy & trauma & phosphate-buffered saline (PBS), and 3) laminectomy+trauma+SCs. Green fluorescent protein (GFP) labelled rPI-SCs were transplanted into the injured rat spinal cord. Their motilities were evaluated with Basso, Beattie and Bresnahan (BBB) Score. After 4-weeks, spinal cord sections were analyzed for GFP labeled SCs and stained for vimentin, $S100{\beta}$, brain derived neurotrophic factor (BDNF), 2',3'-cyclic-nucleotide 3'-phosphodiesterase (CNPase), vascular endothelial growth factor (VEGF) and proinflammatory (interleukin [IL]-6, transforming growth factor $[TGF]-{\beta}$, macrophage inflammatory protein [MIP]-2, myeloperoxidase [MPO]) and anti-inflammatory (IL-1 receptor antagonis) factors. Results : rPI-SCs were revealed to display MSC characteristics and express neural and glial cell markers including BDNF, glial fibrillary acidic protein (GFAP), fibronectin, microtubule associated protein-2a,b (MAP2a,b), ${\beta}3$-tubulin and nestin as well as anti-inflammatory prostaglandin E2 receptor, EP3. The BBB scores showed significant motor recovery in group 3. GFP-labelled cells were localized on the injury site. In addition, decreased proinflammatory factor levels and increased intensity of anti-inflammatory factors were determined. Conclusion : Transplantation of PI-SCs might be an effective strategy to improve functional recovery following spinal cord trauma.

Recovery of Heavy-Metallic Components from a Waste Electro-polishing Solution of 316L Steel by the Solar Cell Electricity (태양전지 전력을 이용한 316L강의 전해연마 폐액 중 중금속 성분의 회수)

  • Kim, Ki-Ho;Jang, Jung-Mok
    • Journal of Surface Science and Engineering
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    • v.42 no.1
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    • pp.53-57
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    • 2009
  • Recovery of heavy-metallic component from a waste solution of factory was undertaken by the solar cell electricity. The solution was obtained from an electrolytic etching process of 316L stainless steel. The electrolysis of the solution for recovery of heavy metallic components was made with platinum plated titanium mesh anode and copper plate cathode. Analysis for the solution and electro-winned materials were made by EDS, XRD and SEM. Iron, chromium, and sulfur components were recovered on the cathode from the solution. Result of EDS analysis for the electro-winned materials revealed that some metal oxide were contained in the recovered material. The recovered materials were expected to have metallic form only by the electrolysis, but metal compounds were contained because of weak solar cell power. Nickel and manganese component in the solution doesn't recovered by this electrolysis process, but they made a sludge with phosphoric acid in the solution.

Olig2-expressing Mesenchymal Stem Cells Enhance Functional Recovery after Contusive Spinal Cord Injury

  • Park, Hwan-Woo;Oh, Soonyi;Lee, Kyung Hee;Lee, Bae Hwan;Chang, Mi-Sook
    • International Journal of Stem Cells
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    • v.11 no.2
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    • pp.177-186
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    • 2018
  • Background and Objectives: Glial scarring and inflammation after spinal cord injury (SCI) interfere with neural regeneration and functional recovery due to the inhibitory microenvironment of the injured spinal cord. Stem cell transplantation can improve functional recovery in experimental models of SCI, but many obstacles to clinical application remain due to concerns regarding the effectiveness and safety of stem cell transplantation for SCI patients. In this study, we investigated the effects of transplantation of human mesenchymal stem cells (hMSCs) that were genetically modified to express Olig2 in a rat model of SCI. Methods: Bone marrow-derived hMSCs were genetically modified to express Olig2 and transplanted one week after the induction of contusive SCI in a rat model. Spinal cords were harvested 7 weeks after transplantation. Results: Transplantation of Olig2-expressing hMSCs significantly improved functional recovery in a rat model of contusive SCI model compared to the control hMSC-transplanted group. Transplantation of Olig2-expressing hMSCs also attenuated glial scar formation in spinal cord lesions. Immunohistochemical analysis showed that transplanted Olig2-expressing hMSCs were partially differentiated into Olig1-positive oligodendrocyte-like cells in spinal cords. Furthermore, NF-M-positive axons were more abundant in the Olig2-expressing hMSC-transplanted group than in the control hMSC-transplanted group. Conclusions: We suggest that Olig2-expressing hMSCs are a safe and optimal cell source for treating SCI.

Comparison of Mechanical Allodynia and Recovery of Locomotion and Bladder Function by Different Parameters of Low Thoracic Spinal Contusion Injury in Rats

  • Carter, Michael W.;Johnson, Kathia M.;Lee, Jun Yeon;Hulsebosch, Claire E.;Gwak, Young Seob
    • The Korean Journal of Pain
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    • v.29 no.2
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    • pp.86-95
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    • 2016
  • Background: The present study was designed to examine the functional recovery following spinal cord injury (SCI) by adjusting the parameters of impact force and dwell-time using the Infinite Horizon (IH) impactor device. Methods: Sprague-Dawley rats (225-240 g) were divided into eight injury groups based on force of injury (Kdyn) and dwell time (seconds), indicated as Force-Dwell time: 150-4, 150-3, 150-2, 150-1, 150-0, 200-0, 90-2 and sham controls, respectively. Results: After T10 SCI, higher injury force produced greater spinal cord displacement (P < 0.05) and showed a significant correlation (r = 0.813) between the displacement and the force (P < 0.05). In neuropathic pain-like behavior, the percent of paw withdrawals scores in the hindpaw for the 150-4, 150-3, 150-2, 150-1 and the 200-0 injury groups were significantly lowered compared with sham controls (P < 0.05). The recovery of locomotion had a significant within-subjects effect of time (P < 0.05) and the 150-0 group had increased recovery compared to other groups (P < 0.05). In addition, the 200-0 and the 90-2 recovered significantly better than all the 150 kdyn impact groups that included a dwell-time (P < 0.05). In recovery of spontaneous bladder function, the 150-4 injury group took significantly longer recovery time whereas the 150-0 and the 90-2 groups had the shortest recovery times. Conclusions: The present study demonstrates SCI parameters optimize development of mechanical allodynia and other pathological outcomes.

ELECTROLYTIC RECOVERY OF PRECIOUS METALS FROM DILUTED SOLUTIONS

  • Roland Kammel;Lieber, Hans-Wilhelm
    • Journal of Surface Science and Engineering
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    • v.14 no.4
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    • pp.209-214
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    • 1981
  • Electrolytic recovery of gold and silver from diluted solutions has at first been applied nearly 100 years ago. Compared with other recovery techniques electrolytic treatment offers several advantages. But to achieve high space-time yields special constructions of electrolytic cells for improved mass transfer are necessary. Ex-tended cathode surface areas and vigorous agitation are important features. Gold and silver recovery by use of newly developed electrolytic cell constructions - rotating tubular bed reactor and impact rod reactor - has been described. Calculations based on industrial application during several years are confirming that electrolytic recovery of precious metalss from diluted solutions has been operated in a very economic way.

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CD40 Co-stimulation Inhibits Sustained BCR-induced $Ca^{2+}$ Signaling in Response to Long-term Antigenic Stimulation of Immature B Cells

  • Nguyen, Yen Hoang;Lee, Ki-Young;Kim, Tae-Jin;Kim, Sung-Joon;Kang, Tong-Mook
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.3
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    • pp.179-187
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    • 2011
  • Regulation of B cell receptor (BCR)-induced $Ca^{2+}$ signaling by CD40 co-stimulation was compared in long-term BCR-stimulated immature (WEHI-231) and mature (Bal-17) B cells. In response to long-term pre-stimulation of immature WEHI-231 cells to ${\alpha}$-IgM antibody (0.5~48 hr), the initial transient decrease in BCR-induced $[Ca^{2+}]_i$ was followed by spontaneous recovery to control level within 24 hr. The recovery of $Ca^{2+}$ signaling in WEHI-231 cells was not due to restoration of internalized receptor but instead to an increase in the levels of $PLC{\gamma}2$ and $IP_3R-3$. CD40 co-stimulation of WEHI-231 cells prevented BCR-induced cell cycle arrest and apoptosis, and it strongly inhibited the recovery of BCR-induced $Ca^{2+}$ signaling. CD40 co-stimulation also enhanced BCR internalization and reduced expression of $PLC{\gamma}2$ and $IP_3R-3$. Pre-treatment of WEHI-231 cells with the antioxidant N-acetyl-L-cysteine (NAC) strongly inhibited CD40-mediated prevention of the recovery of $Ca^{2+}$ signaling. In contrast to immature WEHI-231 cells, identical long-term ${\alpha}$-IgM pre-stimulation of mature Bal-17 cells abolished the increase in BCR-induced $[Ca^{2+}]_i$, regardless of CD40 co-stimulation. These results suggest that CD40-mediated signaling prevents antigen-induced cell cycle arrest and apoptosis of immature B cells through inhibition of sustained BCR-induced $Ca^{2+}$ signaling.

Effects of Ozone Oxidation on the Contact Resistance of DRAM Cell (오존 산화가 DRAM 셀의 콘택 저항에 미치는 영향)

  • 최재승;이승욱;신봉조;박근형;이재봉
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.17 no.2
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    • pp.121-126
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    • 2004
  • In this paper, the effects of the ozone oxidation of the landing polycrystalline silicon on the cell contact resistance of the DRAM device were studied. For this study, the ozone oxidation of the landing polycrystalline silicon layer was performed under various conditions, which was followed by the normal DRAM processes. Then, the cell contact resistance and $t_{WR}$ (write recovery time) of the devices were measured and analyzed. The experimental results showed that the cell contact resistance was more significantly increased for higher temperature of oxidation, longer time of oxidation, and higher concentration of ozone in the oxidation furnace. In addition, the TEM cross-sectional micrographs clearly showed that the oxide layer at the interface between the landing polycrystalline silicon layer and the plug polycrystalline silicon layer was increased by the ozone oxidation. Furthermore, the rate of the device failure due to too large write recovery time was also found to be well correlated with the increase of the cell contact resistance.

Enhancing Extraction Yield of Chlorella Extract by Enzyme Treatment

  • In, Man-Jin;Jang, Jae-Eun;Kim, Dong-Ho
    • Journal of Applied Biological Chemistry
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    • v.50 no.3
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    • pp.132-135
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    • 2007
  • An efficient production method of chlorella extract was developed by enzymatic treatment using cell lytic and proteolytic enzymes. The suitable dosage of Tunicase, a cell lytic enzyme, was found to be 1.0% (w/w). Proteolytic enzymes were screened to obtain high chlorella growth factor (CGF) index, which indicates crude CGF content and solid recovery. Among the seven tested proteases, Esperase, whose optimal dosage was 1.0% (w/w), was selected. By co-treatment using optimal dosages of Tunicase and Esperase, the highest CGF index and solid recovery were obtained. The CGF index and solid recovery of co-treatment were remarkably enhanced by 250 ($4.36{\rightarrow}15.21$) and 220% ($12.65%{\rightarrow}40.15%$), respectively, than those of the non-treated extracts.