• Title/Summary/Keyword: tissue-engineering

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Antibody Production in Plant Cell Cultures

  • Lee, James M.
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.67-78
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    • 1995
  • Monoclonal antibodies (MoAbs) are a highly diversified class of proteins with major research and commercial applications such as diagnostics and therapeutics. Currently, the dominant method for producing MoAbs is through the hybridoma technique. However, this technique is slow, tedious, labor intensive, and expensive. The production of MoAbs in cultured transgenic plant cells can offer some advantages over that in the over that in the mammalian systems. The media to cultivate plant cells are well defined and inexpensive. Contamination by bacteria or fungi is easily monitored in plant tissue cultures. Furthermore, these contaminants are usually not potent pathogens to human beings. In our interdisciplinary research efforts, heavy chain monoclonal antibody (HC MAb) was inserted into Ti plasmid vector and transferred into A. tumefaciens for the transformation in tobacco cells. It was found that 76% of the transformants produced HC MAb. The presence of HC MAb in the cell membrane fraction indicated that the signal peptide was functional and efficient. The change of the HC MAb concentration during a batch culture followed a similar trend as dry cell concentration, indicating that the production of HC MAb was growth related. The long-term repeated subcultures of 11 cell lines showed that there was no obvious trend of neither the decrease nor the increase of the productivity with the repeated subcultures.

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Molecular Identification and Expression of Myosin Light Chain in Shortspine Spurdog (Squalus mitsukurii)

  • Kim, Soo Cheol;Sumi, Kanij Rukshana;Sharker, Md Rajib;Kho, Kang Hee
    • Journal of Marine Life Science
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    • v.3 no.1
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    • pp.1-8
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    • 2018
  • Myosin is considered as the vital motor protein in vertebrates and invertebrates. Our present study was conducted to decipher the occurrence of myosin in dog fish (Squalus mitsukurii). We isolated one clone containing 979 bp cDNA sequence, which consisted of a complete coding sequence of 453 bp and a deduced amino acid sequence of 150 amino acids from the open reading frame with molecular weight, isoelectric point and aliphatic index are 16.72 Kda, 4.49 and 78.00, respectively. It contained 428 bp long 3' UTR with single potential polyadenylation signals (AATAAA). The predicted EF CA2+ binding domains were identified in residue 6-41, 83-118 and 133-150. A BLAST search indicates this protein exhibits a strong similarity to whale shark (Rhincodon typus) MLC3 (91% identical) and also house mouse (Mus musculus) MLC isoform 3f (81% identical). Phylogenetic analysis revealed that this protein is a MLC 3 isoform like protein. This protein also demonstrates highly conserved region with other myosin proteins. Homology modeling of S. mitsukuri was performed using crystal structure of Gallus gallus skeletal muscle myosin II based on high similarity. Reverse transcription-polymerase chain reaction (PCR), quantitative PCR results exhibits dogfish myosin protein is highly expressed in muscle tissue.

Analysis of UCP1 Expression in Rainbow Trout Gonadal Cell Line RTG-2 Indicates its Marginal Response to Adipogenic Inducers Compared to Mammalian Cell Lines

  • Sang-Eun Nam;Young-Joo Yun;Jae-Sung Rhee;Hyoung Sook Park
    • Journal of Marine Life Science
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    • v.8 no.2
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    • pp.186-189
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    • 2023
  • Uncoupling protein 1 (UCP1) is a unique mitochondrial membranous protein expressed in brown adipose tissue (BAT) in mammals. While its expression in response to cold temperatures and adipogenic inducers is well-characterized in mammals and human infants, the molecular characterization and expression of UCP1 in fish remain unexplored. To address this gap, we analyzed UCP1 expression in response to adipogenic inducers in a fish cell line, rainbow trout gonadal cells (RTG-2), and compared it with UCP1 expression in three mammalian preadipocytes, 3T3-L1, T37i, and WT1 exposed to the Peroxisome proliferator-activated receptor gamma (PPARγ) agonists, rosiglitazone (Rosi). In mammalian preadipocytes, UCP1 protein was highly expressed by Rosi, with an induction of adipogenesis observed in a time-dependent manner. This suggests that UCP1 plays a significant role in adipogenesis in mammals. However, RTG-2 cells showed no response to adipogenic inducers and exhibited only marginal expressions of UCP1. These results imply that RTG-2 cells may lack crucial responsive mechanisms to adipogenic signals or that the adipogenic response is regulated by other mechanisms. Further studies are needed to confirm these phenomena in fish preadipocytes when an appropriate cell line is established in future research.

Effect of Blood Contamination on Vickers Microhardness and Surface Morphology of Mineral Trioxide Aggregate

  • Jaehyun Seung;Seong-Jin Shin;Byounghwa Kim;Ji-Myung Bae;Jiyoung Ra
    • Journal of the korean academy of Pediatric Dentistry
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    • v.51 no.2
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    • pp.165-175
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    • 2024
  • This study aimed to investigate the effects of blood contamination on the Vickers hardness and the surface morphology of premixed MTA and compare them with the effects on conventional MTA. The Vickers microhardness of Endocem MTA Premixed Regular (EP) and ProRoot MTA (PM) was assessed after immersion in fetal bovine serum (FBS) and saline. Stem cells from human exfoliated deciduous teeth (SHED) were seeded on MTA after immersion in FBS, saline, and deionized water (DW). Cell adhesion patterns and surface morphology were visualized via scanning electron microscopy (SEM). The surface microhardness of EP and PM in FBS was lower than in saline. However, short-term exposure of PM to FBS did not reduce the microhardness compared to saline. Angular crystals formed in water, while rounded crystals with more air voids appeared in FBS. Favorable SHED attachment occurred in all groups. Overall, the surface hardness of EP and PM decreased after FBS exposure, although PM was less influenced. We suggest minimizing the amount of bleeding when using MTA clinically; nevertheless, PM remains an option with more expected blood contamination than EP. In summary, exposure to FBS decreased mechanical performance but allowed cell adhesion for both MTAs, with PM being more resistant to these changes.

IL-15 in T-Cell Responses and Immunopathogenesis

  • Hoyoung Lee;Su-Hyung Park;Eui-Cheol Shin
    • IMMUNE NETWORK
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    • v.24 no.1
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    • pp.11.1-11.18
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    • 2024
  • IL-15 belongs to the common gamma chain cytokine family and has pleiotropic immunological functions. IL-15 is a homeostatic cytokine essential for the development and maintenance of NK cells and memory CD8+ T cells. In addition, IL-15 plays a critical role in the activation, effector functions, tissue residency, and senescence of CD8+ T cells. IL-15 also activates virtual memory T cells, mucosal-associated invariant T cells and γδ T cells. Recently, IL-15 has been highlighted as a major trigger of TCR-independent activation of T cells. This mechanism is involved in T cell-mediated immunopathogenesis in diverse diseases, including viral infections and chronic inflammatory diseases. Deeper understanding of IL-15-mediated T-cell responses and their underlying mechanisms could optimize therapeutic strategies to ameliorate host injury by T cell-mediated immunopathogenesis. This review highlights recent advancements in comprehending the role of IL-15 in relation to T cell responses and immunopathogenesis under various host conditions.

Alleviating Effects of Nitric Oxide on Cadmium Toxicity in White Poplar (Populus alba)

  • Semsettin Kulac;Yakup Cikili;Halil Samet;Ertugrul Filiz
    • Journal of Forest and Environmental Science
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    • v.40 no.1
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    • pp.43-52
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    • 2024
  • Cadmium (Cd) is non-essential heavy metal that negatively affects plant metabolism. Nitric oxide (NO) is an increasingly important molecule for plant metabolism that makes signaling. In this study, it was aimed to investigate the alleviating effect of sodium nitroprusside (SNP) application as NO donor in white poplar (Populus alba) under Cd stress conditions. SNP and without SNP treatments increased the Cd accumulation in root tissue. While photosynthetic pigments (Chl a, Chl b, Chl a+b, and carotenoid) content decreased by only Cd application, SNP+Cd application decreased the rate of photosynthetic pigments reduction. When the results of Cd and Cd+SNP applications were evaluated for mineral (Fe, Zn, Mn and Cu) uptake, it was found that the positive effect of SNP was heterogeneously affected. Depending on SNP application, it was found that malondialdehyde (MDA) amount decreased in leaf in 100 µM Cd applications while hydrogen peroxide (H2O2) amount decreased in 100 and 500 µM Cd applications. When antioxidant enzyme activities were examined, it was found that catalase (CAT) and ascorbate peroxidase (APX) enzyme activities increased with 100 µM SNP applications under all Cd applications. As a result, it was found that SNP application under Cd stress generally supports physiological processes positively in white poplar, suggesting that NO molecule plays important alleviating roles in plant metabolism.

Tumor Stroma as a Therapeutic Target for Pancreatic Ductal Adenocarcinoma

  • Dae Ui Lee;Beom Seok Han;Kyung Hee Jung;Soon-Sun Hong
    • Biomolecules & Therapeutics
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    • v.32 no.3
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    • pp.281-290
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    • 2024
  • Pancreatic ductal adenocarcinoma (PDAC) has a poor prognosis owing to its desmoplastic stroma. Therefore, therapeutic strategies targeting this tumor stroma should be developed. In this study, we describe the heterogeneity of cancer-associated fibroblasts (CAFs) and their diverse roles in the progression, immune evasion, and resistance to treatment of PDAC. We subclassified the spatial distribution and functional activity of CAFs to highlight their effects on prognosis and drug delivery. Extracellular matrix components such as collagen and hyaluronan are described for their roles in tumor behavior and treatment outcomes, implying their potential as therapeutic targets. We also discussed the roles of extracellular matrix (ECM) including matrix metalloproteinases and tissue inhibitors in PDAC progression. Finally, we explored the role of the adaptive and innate immune systems in shaping the PDAC microenvironment and potential therapeutic strategies, with a focus on immune cell subsets, cytokines, and immunosuppressive mechanisms. These insights provide a comprehensive understanding of PDAC and pave the way for the development of prognostic markers and therapeutic interventions.

Effect of Demineralized Bone Particle Gel Penetrated into Poly(lactic-co-glycolic acid) Scaffold on the Regeneration of Chondrocyte: In Vivo Experiment (PLGA 다공성 지지체에 함침시킨 DBP젤의 연골재생 효과: In Vivo 실험)

  • Lee, Yun Mi;Shim, Cho Rok;Lee, Yujung;Kim, Ha Neul;Jo, Sun A;Song, Jeong Eun;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.36 no.6
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    • pp.789-794
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    • 2012
  • Poly(lactic-co-glycolic acid) (PLGA) has been most widely used due to its advantages such as good biodegradability, controllable rate of degradation and metabolizable degradation products. We manufactured composite scaffolds of PLGA scaffold penetrated DBP gel (PLGA/DBP gel) by a simple method, solvent casting/salt leaching prep of PLGA scaffolds and subsequent soaking in DBP gel. Chondrocytes were seeded on the PLGA/DBP gel. The mechanical strength of scaffold, histology (H&E, Safranin-O, Alcian-blue) and immunohistochemistry (collagen type I, collagen type II) were performed to elucidate in vitro and in vivo cartilage-specific extracellular matrices. It was better to keep the characteristic of chondrocytes in the PLGA/DBP gel scaffolds than that PLGA scaffolds. This study suggests that PLGA/DBP gel scaffold may serve as a potential cell delivery vehicle and a structural basis for in vivo tissue engineered cartilage.

Molecular Cloning and Characterization of a Novel Stem-specific Gene from Camptotheca acuminata

  • Pi, Yan;Liao, Zhihua;Chai, Yourong;Zeng, Hainian;Wang, Peng;Gong, Yifu;Pang, Yongzhen;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.39 no.1
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    • pp.68-75
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    • 2006
  • In higher plants, P450s participate in the biosynthesis of many important secondary metabolites. Here we reported for the first time the isolation of a new cytochrome P450 cDNA that expressed in a stem-specific manner from Camptotheca acuminata (designated as CaSS), a native medicinal plant species in China, using RACE-PCR. The full-length cDNA of CaSS was 1735 bp long containing a 1530 bp open reading frame (ORF) encoding a polypeptide of 509 amino acids. Bioinformatic analysis revealed that CASS contained a heme-binding domain PFGXGRRXCX and showed homology to other plant cytochrome P450 monooxygenases and hydroxylases. Southern blotting analysis revealed that there was only one copy of the CaSS present in the genome of Camptotheca acuminata. Northern blotting analysis revealed that CaSS expressed, in a tissue-specific manner, highly in stem and lowly in root, leaf and flower. Our study suggests that CaSS is likely to be involved in the phenylpropanoid pathway.

Effects of PLGA/Fibrin Scaffolds on Attachment and Proliferation of Costal Cartilage Cells (PLGA/피브린 지지체가 늑연골 세포의 부착과 성장에 미치는 영향)

  • Song, Jeong Eun;Lee, Yujung;Lee, Yun Me;Cho, Sun Ah;Jang, Ji Eun;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.37 no.2
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    • pp.141-147
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    • 2013
  • Poly(lactide-co-glycolic acid) (PLGA) has been widely used in the drug delivery and tissue engineering applications because of its good mechanical strength and biodegradation profile. However, cell attachment to the scaffold is low compared with that on fibrin although cells can be attached to the polymer surface. In this study, PLGA scaffolds were soaked in cells-fibrin suspension and polymerized with dropping fibrinogen-thrombin solution. Cellular proliferation activity was observed in PLGA/fibrin-seeded costal cartilage cells (CC) on 1, 3, and 7 days using the MTT assay and SEM. The effects of fibrin on the extracellular matrix (ECM) formation were evaluated using CC cell-seeded PLGA/fibrin scaffolds. The PLGA/fibrin scaffolds elicited more production of glycosaminoglycan (GAG) and collagen than the PLGA scaffold. In this study, fibrin incorporated PLGA scaffolds were prepared to evaluate the effects of fibrin on the cell attachment and proliferation in vitro and in vivo. In this result, we confirmed that proliferation of cells in PLGA/fibrin scaffolds were better than in PLGA scaffolds. The PLGA/fibrin scaffolds provide suitable environment for growth and proliferation of costal cartilage cells.