• Title/Summary/Keyword: in vitro Differentiation

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Effects of Cirsium setidens nakai on In Vitro Growth and Osteogenic Differentiation of Human Bone-Derived Mesenchymal Stem Cells

  • Kim, Hye-Been;Cheong, Kyu Min;Seo, Yu Ri;Lim, Ki-Taek
    • Proceedings of the Korean Society for Agricultural Machinery Conference
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    • 2017.04a
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    • pp.109-109
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    • 2017
  • Cirsium setidens nakai belonging to cirsium has been reported to have various physiological activities including anticancer activity because it contains polyphenols, dietary fiber, minerals and vitamins. Despite these positive positive efficacies, however, no studies have studied cirsium setidens nakai products as biomaterials such as cellular metabolism and bone formation. Thus, the aim of this study was evaluate of osteogenesis differentiation a natural material extracted from cirsium setidens nakai. The natural materials in this studys in this studywere created by 40% ethanol extraction process and then dried. FabricatedFabricatedpowders were added to a medium at various concentrations (0.01, 0.05, 0.1, 0.2, and $0.25{\mu}g/mL$), and pure medium was used as a control. The natural material caused positive increases in cell metabolic activity and mineralized bone formation without cytotoxicity. In addition, we observed higher expression of genes such as ALP, BSP, Runx2 and COL1 in cirsium setidens nakai treatment cells. As a result, this study produced and investigated cirsium setidens nakai extracts and the natural materials showed potential biomaterials. In this research indicated that the cirsium setidens nakai extracts might have promising applications in areas of agricultural, biological and food engineering as a biomaterial.

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CD43 Expression Regulated by IL-12 Signaling Is Associated with Survival of CD8 T Cells

  • Lee, Jee-Boong;Chang, Jun
    • IMMUNE NETWORK
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    • v.10 no.5
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    • pp.153-163
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    • 2010
  • Background: In addition to TCR and costimulatory signals, cytokine signals are required for the differentiation of activated CD8 T cells into memory T cells and their survival. Previously, we have shown that IL-12 priming during initial antigenic stimulation significantly enhanced the survival of activated CD8 T cells and increased the memory cell population. In the present study, we analyzed the mechanisms by which IL-12 priming contributes to activation and survival of CD8 T cells. Methods: We observed dramatically decreased expression of CD43 in activated CD8 T cells by IL-12 priming. We purified $CD43^{lo}$ and $CD43^{hi}$ cells after IL-12 priming and analyzed the function and survival of each population both in vivo and in vitro. Results: Compared to $CD43^{hi}$ effector cells, $CD43^{lo}$ effector CD8 T cells exhibited reduced cytolytic activity and lower granzyme B expression but showed increased survival. $CD43^{lo}$ effector CD8 T cells also showed increased in vivo expansion after adoptive transfer and antigen challenge. The enhanced survival of $CD43^{lo}$ CD8 T cells was also partly associated with CD62L expression. Conclusion: We suggest that CD43 expression regulated by IL-12 priming plays an important role in differentiation and survival of CD8 T cells.

Evaluation of Skin Sebosuppression by Components of Total Green Tea (Camellia sinensis) Extracts

  • Kim, Jeong-Kee;Shin, Hyun-Jung;Lee, Byeong-Gon;Lee, Sang-Jun
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.464-469
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    • 2008
  • In human beings, it is known that there is a correlation between the occurrence of acne and the ability to suppress sebum. Sebosuppression may be related to the inhibition of sebocyte proliferation, differentiation, and lipogenesis in sebaceous glands. To investigate the skin sebosuppressive activity of green tea extract, the in vivo effects of its flavonoid compounds on the androgen-dependent stimulation of pigmented macules in hamsters and performed in vitro experiments with human primary sebocytes were examined. Our results imply a dual activity of skin sebosuppression by green tea flavonoids; some catechins including epigallocatechin-3-gallate (EGCG) and gallocatechin-3-gallate (GCG) may reduce the differentiation of sebocytes by inhibiting PPAR-${\gamma}1$ mRNA expression, whereas some flavonol glycosides including kaempferol may inhibit lipogenesis in sebaceous glands by decreasing levels of the mature form of sterol-sensitive response elements binding protein-1c (SREBP-1c). Therefore, green tea is a potentially effective material for use in the development of health foods or cosmetics for skin sebosuppression.

Activation of CREB by PKA Promotes the Chondrogeneic Differentiation of Chick Limb Bud Mesenchymal Cells

  • Kim, Kook-Hee;Lee, Young-Sup
    • Animal cells and systems
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    • v.13 no.3
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    • pp.289-295
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    • 2009
  • Cyclic AMP-mediated signaling pathways regulate a number of cellular functions. In this study, we examined the regulatory role of cAMP signaling pathways in chondrogenesis of chick limb bud mesenchymal cells in vitro. Forskolin, which increases cellular cAMP levels by the activation of adenylate cyclase, enhanced chondrogenic differentiation. Inhibition of PKA with specific inhibitors (H89 or KT5720) blocked pre-cartilage condensation stage, indicating that chondrogenesis is regulated by the increase in cellular cAMP level and subsequent activation of PKA. Downstream signaling pathway of PKA leading to gene expression was investigated by examination of several nuclear transcription factors. Forskolin treatment increased transcription level for a cartilage-specific marker gene Sox9. However, inhibition of PKA with H89 led to restore expression of Sox9, indicating PKA activity was required to regulate the expression of Sox9 in chondrogenesis. In addition, CREB was highly phosphorylated at early stage of mesenchyme culture, and followed by progressive dephosphorylation. CBP and ATF, another CRE related proteins were transiently expressed at the early stage of chondrogenesis with a pattern similar to CREB phosphorylation. Electrophoretic mobility shift assays confirmed that the binding activity of CREB to the CRE is closely correlated to the phosphorylation pattern of CREB. Therefore, cAMP-mediated signal transduction to nuclear events for the induction of genes appeared to be required at the early stage of chick limb bud chondrogenesis.

Cloning and characterization of polyA- RNA transcripts encoded by activated B1-like retrotransposons in mouse erythroleukemia MEL cells exposed to methylation inhibitors

  • Tezias, Sotirios S.;Tsiftsoglou, Asterios S.;Amanatiadou, Elsa P.;Vizirianakis, Ioannis S.
    • BMB Reports
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    • v.45 no.2
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    • pp.126-131
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    • 2012
  • We have previously identified a DNA silent region located downstream of the 3'-end of the ${\beta}^{major}$ globin gene (designated B1-559) that contains a B1 retrotransposon, consensus binding sites for erythroid specific transcription factors and shares the capacity to act as promoter in hematopoietic cells interacting with ${\beta}$-globin gene LCR sequences in vitro. In this study, we have cloned four new non-polyA RNA transcripts being detected upon blockade of murine erythroleukemia (MEL) cell differentiation to erythroid maturation by methylation inhibitors and demonstrated that two of them share high structural homology with sequences of B1 element found within the B1-559 region. Although it is not clear yet whether and how these RNAs interfere with induction of erythroid maturation, these data provide evidence for the first time showing that methylation inhibitors can activate silent repetitive DNA sequences in MEL cells and may have implications in cancer chemotherapy using demethylating drugs as antineoplastic agents.

Protocadherin-7 contributes to maintenance of bone homeostasis through regulation of osteoclast multinucleation

  • Kim, Hyunsoo;Takegahara, Noriko;Walsh, Matthew C.;Ueda, Jun;Fujihara, Yoshitaka;Ikawa, Masahito;Choi, Yongwon
    • BMB Reports
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    • v.53 no.9
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    • pp.472-477
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    • 2020
  • Osteoclasts are hematopoietic-derived cells that resorb bone. They are required to maintain proper bone homeostasis and skeletal strength. Although osteoclast differentiation depends on receptor activator of NF-κB ligand (RANKL) stimulation, additional molecules further contribute to osteoclast maturation. Here, we demonstrate that protocadherin-7 (Pcdh7) regulates formation of multinucleated osteoclasts and contributes to maintenance of bone homeostasis. We found that Pcdh7 expression is induced by RANKL stimulation, and that RNAi-mediated knockdown of Pcdh7 resulted in impaired formation of osteoclasts. We generated Pcdh7-deficient mice and found increased bone mass due to decreased bone resorption but without any defect in bone formation. Using an in vitro culture system, it was revealed that formation of multinucleated osteoclasts is impaired in Pcdh7-deficient cultures, while no apparent defects were observed in differentiation and function of Pcdh7-deficient osteoblasts. Taken together, these results reveal an osteoclast cell-intrinsic role for Pcdh7 in maintaining bone homeostasis.

Modified Adenovirus Mediated Gene Transfer to Neuronal Precursor Cells (Transferrine peptide ligand로 개량된 아데노바이러스를 이용한 신경전구세포로의 유전자 전달 효율 조사)

  • Joung, In-Sil
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.73-76
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    • 2006
  • Neuronal precursor cells may provide for cell replacement or gene delivery vehicles in neurodegenerative disease therapy. One impediment to treating neuronal diseases is finding ways to introduce genes into neurons effectively. It is shown here that fiber-modified adenovirus vector delivered gene to neuronal precursor as well as differentiated neuronal cells more efficiently than first-generation adenoviral vector. Moreover, fiber-modified adenoviral vector transduced precursor cells retained the potential for differentiation into neurons and glia in vitro. These results show the potential of modified adenoviral vector in the improved gene delivery to neurons in direct gene therapy protocols. In addition it holds promise for the use of genetically manipulated stem cells for the therapy of neuronal diseases.

Different Pattern of p27kip1 and p21cip1 Expression Following Ex Vivo Activation of CD8+ T Lymphocytes

  • Kim, Sung-Jin;Lee, Hyeon-Woo
    • Biomolecules & Therapeutics
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    • v.15 no.4
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    • pp.218-223
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    • 2007
  • T cell proliferation is a pivotal to an effective immune response. Cyclin-dependent kinase (cdk) inhibitor, $p27^{kip1}$ is degraded to initiate T cell expansion. In this study, we show that although the expression of $p27^{kip1}$ protein was down-regulated, that of $p21^{cip1}$, another cdk inhibitor, was up-regulated in $CD8^+$ T cells following in vitro stimulation. Ex vivo gB antigen-stimulation following HSV immunization increased $p21^{cip1}$ positive cells that co-expressed IFN-$\gamma$. Moreover, $p21^{cip1}$ was co-expressed with IFN-${\gamma}$ in E7 antigen-stimulated $CD8^+$ T cells, whereas $p27^{kip1}$ was not. Our findings imply a role of $p21^{cip1}$ proteins in antigen-induced effector $CD8^+$ T cells differentiation in vivo.

Investigation of Novel Pharmacological Action of Arctii Fructus and its Compound

  • Hong, Seung-Heon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.9-9
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    • 2018
  • Arctii Fructus (AF), which contains arctigenin (ARC) as a major constituent, is traditionally used as an anti-inflammatory medicine to treat inflammatory sore throat. Although several studies have proven its anti-inflammatory effects, there have been no reports on its use in inflammation related disorders such as obesity, cancer metastasis, and allergic responses. This study investigated the anti-obesity effect and anti-metastasis effect of AF and ARC. AF and ARC inhibited weight gain by reducing the mass of white adipose tissue in high fat diet (HFD)-induced obese mice. Serum cholesterol levels were also improved by AF and ARC. In in vitro experiments, AF and ARC decreased differentiation of white adipocytes. Furthermore, AF induced differentiation of brown adipocytes, which are able to consume surplus energy through non-shivering thermogenesis. Also, AF and ARC inhibited colon cancer and lung metastasis of colon cancer. They suppressed not only colorectal cancer cell progression by inhibiting cell growth, but also prohibited lung metastasis by regulating epithelial-mesenchymal transition (EMT), migration, and the invasion. These effects were confirmed in an experimental metastasis mouse model. In addition, AF and ARC inhibited mast cell mediated allergic responses. Collectively, our study suggests that AF and ARC might show inhibitory effects on inflammation related diseases, including obesity, cancer, cancer metastasis, and allergic responses.

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Anti-adipocyte differentiation activity and flavonoid content determination by HPLC/UV analysis of tree sprouts

  • Kim, Juree;Jang, Taewon;Kim, Ji Hyun;Shin, Hanna;Park, Jaeho;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • v.64 no.3
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    • pp.269-275
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    • 2021
  • The in vitro anti-obesity activity of 12 species of tree sprouts in differentiated 3T3-L1 cells and the mechanisms underlying their activity were evaluated. (+)-Catechin and quercetin concentrations in the sprouts were analyzed by HPLC/UV at 270 and 254 nm, respectively. Euonymus alatus (EAT) and Fraxinus mandschuria (FMS) extracts at doses of 50 and 100 ㎍/mL inhibited the accumulation of lipid droplets in differentiated 3T3-L1 cells. Moreover, EAT and FMS downregulated the expression of the CCAAT/enhancer-binding protein-α, adipogenesis-related proteins peroxisome proliferator-activated receptor-γ, and adipocyte P-2α in differentiated 3T3-L1 cells. Tree sprouts with an abundant flavonoid content exerted the highest anti-obesity activity. Concentrations of total flavonoids were the highest in FMS (24.281 mg/g DW) sprouts. These findings could be used to develop health-promoting functional foods or supplements derived from tree sprouts.