• Title/Summary/Keyword: mature cell

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Low-Intensity Pulsed Ultrasound Promotes BMP9 Induced Osteoblastic Differentiation in Rat Dedifferentiated Fat Cells

  • Fumiaki Setoguchi;Kotaro Sena;Kazuyuki Noguchi
    • International Journal of Stem Cells
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    • v.16 no.4
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    • pp.406-414
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    • 2023
  • Dedifferentiated fat cells (DFATs) isolated from mature adipocytes have a multilineage differentiation capacity similar to mesenchymal stem cells and are considered as promising source of cells for tissue engineering. Bone morphogenetic protein 9 (BMP9) and low-intensity pulsed ultrasound (LIPUS) have been reported to stimulate bone formation both in vitro and in vivo. However, the combined effect of BMP9 and LIPUS on osteoblastic differentiation of DFATs has not been studied. After preparing DFATs from mature adipose tissue from rats, DFATs were treated with different doses of BMP9 and/or LIPUS. The effects on osteoblastic differentiation were assessed by changes in alkaline phosphatase (ALP) activity, mineralization/calcium deposition, and expression of bone related genes; Runx2, osterix, osteopontin. No significant differences for ALP activity, mineralization deposition, as well as expression for bone related genes were observed by LIPUS treatment alone while treatment with BMP9 induced osteoblastic differentiation of DFATs in a dose dependent manner. Further, co-treatment with BMP9 and LIPUS significantly increased osteoblastic differentiation of DFATs compared to those treated with BMP9 alone. In addition, upregulation for BMP9-receptor genes was observed by LIPUS treatment. Indomethacin, an inhibitor of prostaglandin synthesis, significantly inhibited the synergistic effect of BMP9 and LIPUS co-stimulation on osteoblastic differentiation of DFATs. LIPUS promotes BMP9 induced osteoblastic differentiation of DFATs in vitro and prostaglandins may be involved in this mechanism.

The Effector Functions of Mature T Lymphocytes Are Impaired in Transgenic Mice Expressing the SH2 Domain of TSAd/Lad

  • Choi, Youngbong;Park, Eunkyung;Ahn, Eunseon;Park, Inyoung;Yun, Yungdae
    • Molecules and Cells
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    • v.28 no.3
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    • pp.183-188
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    • 2009
  • TSAd/Lad is a T cell adaptor molecule involved in $p56^{lck}$-mediated T cell activation. To investigate the functions of TSAd in T cells, we generated transgenic (TG) mice expressing the SH2 domain of TSAd (TSAd-SH2) under the control of the $p56^{lck}$ proximal promoter. In T cells from TSAd-SH2 TG mice, T cell receptor (TCR)-mediated early signaling events, such as $Ca^{2+}$ flux and ERK activation, were normal; however, late activation events, such as IL-2 production and proliferation, were significantly reduced. Moreover, TCR-induced cell adhesion to extracellular matrix (ECM) proteins and migration through ECM proteins were defective in T cells from TSAd-SH2 TG mice. Furthermore, the contact hypersensitivity (CHS) reaction, an inflammatory response mainly mediated by T helper 1 (Th1) cells, was inhibited in TSAd-SH2 TG mice. Taken together, these results show that TSAd, particularly the SH2 domain of TSAd, is essential for the effector functions of T cells.

The Role of High-throughput Transcriptome Analysis in Metabolic Engineering

  • Jewett, Michael C.;Oliveira, Ana Paula;Patil, Kiran Raosaheb;Nielsen, Jens
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.5
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    • pp.385-399
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    • 2005
  • The phenotypic response of a cell results from a well orchestrated web of complex interactions which propagate from the genetic architecture through the metabolic flux network. To rationally design cell factories which carry out specific functional objectives by controlling this hierarchical system is a challenge. Transcriptome analysis, the most mature high-throughput measurement technology, has been readily applied In strain improvement programs in an attempt to Identify genes involved in expressing a given phenotype. Unfortunately, while differentially expressed genes may provide targets for metabolic engineering, phenotypic responses are often not directly linked to transcriptional patterns, This limits the application of genome-wide transcriptional analysis for the design of cell factories. However, improved tools for integrating transcriptional data with other high-throughput measurements and known biological interactions are emerging. These tools hold significant promise for providing the framework to comprehensively dissect the regulatory mechanisms that identify the cellular control mechanisms and lead to more effective strategies to rewire the cellular control elements for metabolic engineering.

High-Level Expression of Recombinant Human Bone Morphogenetic Protein-4 in Chinese Hamster Ovary Cells

  • PARK JUNHO;YU SUNGRYUL;YOON JAESEUNG;BAEK KWANGHEE
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1397-1401
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    • 2005
  • Bone morphogenetic protein-4 (BMP-4) is a signaling homodimeric molecule that acts as a morphogen to influence cell fate in a concentration-dependent manner. The limited supply of a pure preparation of BMP-4, due to very low level of their expression in vivo, makes it difficult not only to study the biological activities of BMPs, but also to use them as a clinical tool. For a large-scale production of BMP-4, human BMP-4 cDNA was expressed in Chinese hamster ovary (CHO) cells by a recently development vector system, which confers position-independent stable expression of the foreign genes. The CHO cell line expressing recombinant human BMP-4 (rhBMP-4) at the level of $7\;{\mu}g/ml$ could be obtained after stepwise selection with methotrexate. This level of expression is about 70 times higher than those previously reported. The partially processed form of BMP-4 as well as mature form could be detected, when the aliquots of culture media were analyzed by Western blot. The glycosylation pattern and biological activity of the rhBMP-4 were determined by glycosidase treatment and the induction rate of alkaline phosphatase in mouse osteoblastic cells.

Characteristics of chain-forming cells in Gyrodinium impudicum using fluorescent ConA

  • Park, Eun-Ju;Kim, Gwang-Hoon;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • v.34 no.4
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    • pp.214-219
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    • 1999
  • Cells of the dinoflagellate Gyrodinium impudicum form characteristic chains, which are associated with sugar accumulated on the cell surface. To resolve the relationship between chain-formation and cell surface sugar accumulation, confocal microscopy was used to observe sugar accumulation points in the vegetative cells and long chain-forming cells of G. impudicum cells treated with fluorescent-tagged ConA. In the stationary and exponential phases, both vegetative cells and chain-forming cells were similar to each other in fluorescent intensity. There was no evidence that chain-forming cells had a specific location for sugar accumulation on the cell surface. Most of the cells formed 2-cell chains one day after inoculation, but longer chains consisting of 4-8 cells increased markedly in 4day and 8 day cultures. Gyrodinium impudicum chains usually consist of more than four cells, and chains of 8 or even 16 cells can be observed in mature cultures. Temperature played an importantrole in chain-formation of the cells, threshold temperature for the development of long chain-formation was 19 $^{\circ}$C.

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A Case of Nasal Diffuse Large B-Cell Lymphoma in a Cat

  • Kim, Seong-Jun;Kim, Jun-Young;Oh, Dong-Keun;Cho, Jun-Ho;Park, Hee-Myung;Kang, Min-Hee
    • Journal of Veterinary Clinics
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    • v.35 no.4
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    • pp.141-143
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    • 2018
  • A 6-year-old spayed female Turkish Angora cat presented with sneezing, nasal discharge, and decreased appetite lasting for 21 days. Skull radiography revealed slightly increased density of soft tissue in the left nasal cavity. Computed tomography (CT) scan revealed an extensive mass with nasal septum destruction and moderate contrast enhancement in the left nasal cavity. After surgical biopsy, histopathological examination confirmed that the mass was an infiltrative round cell neoplasm, composed of sheets of large neoplastic cells. Immunohistochemical analysis revealed that most of the neoplastic cells were strongly positive for CD79a and weakly positive for PAX5. Additionally, numerous mature lymphocytes were found to be positive for CD3. This is the first reported case of nasal diffuse large B-cell lymphoma (DLBCL) in a Turkish Angora cat in Korea.

The Role of Dendritic Cells in Central Tolerance

  • Oh, Jaehak;Shin, Jeoung-Sook
    • IMMUNE NETWORK
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    • v.15 no.3
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    • pp.111-120
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    • 2015
  • Dendritic cells (DCs) play a significant role in establishing self-tolerance through their ability to present self-antigens to developing T cells in the thymus. DCs are predominantly localized in the medullary region of thymus and present a broad range of self-antigens, which include tissue-restricted antigens expressed and transferred from medullary thymic epithelial cells, circulating antigens directly captured by thymic DCs through coticomedullary junction blood vessels, and peripheral tissue antigens captured and transported by peripheral tissue DCs homing to the thymus. When antigen-presenting DCs make a high affinity interaction with antigen-specific thymocytes, this interaction drives the interacting thymocytes to death, a process often referred to as negative selection, which fundamentally blocks the self-reactive thymocytes from differentiating into mature T cells. Alternatively, the interacting thymocytes differentiate into the regulatory T (Treg) cells, a distinct T cell subset with potent immune suppressive activities. The specific mechanisms by which thymic DCs differentiate Treg cells have been proposed by several laboratories. Here, we review the literatures that elucidate the contribution of thymic DCs to negative selection and Treg cell differentiation, and discusses its potential mechanisms and future directions.

Alternation in the Cell-mediated Immune Reaction by Bisphenol A Exposure in vitro (In vitro에서 Bisphenol A 노출에 의한 세포성 면역반응의 변화)

  • Pyo Myoung-Yun
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.92-96
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    • 2005
  • In order to investigate the effects of bisphenol A (BPA) on cell mediated immune reaction in vitro we examined the allogenic mixed lymphocyte reaction (MLR), splenocytes proliferation (SP) to T cell mitogens and IFN-${\gamma}\;production$. Splenocytes of Balb/c mice ($1.5{\times}10^5$ cells/well) were co-cultured with different numbers of mitomycin C-treated mature dentritic cells (DCs) in presence of BPA (25, 50, 100 ${\mu}M$) and $[^{3}H]$thymidine incorporation (cpm) was measured by scintilation counting. Splenocytes ($2{\times}10^6$ cells/well) were cultured with mitogens, Con A ($2\;{\mu}g/ml$), PHA ($5\;{\mu}g/ml$) and IL-2 ($0.1\;{\mu}g/ml$), or PMA ($5\;{\mu}g/ml$) and INO ($1\;{\mu}g/ml$) in presence of BPA (1, 10, 25, 50, 100 ${\mu}M$) and SP was assessed by MTT assay. $IFN-{\gamma}$ levels in culture supernant were determined by ELISA. At low concentration, BPA slightly increased MLR, SP and $IFN-{\gamma}$ levels, but at higher concentration it showed significant inhibitory effects on these immunological parameters. These results indicate that BPA is able to alternate cell-mediated immune reaction.

Ultrastructure of the Gill of the Parrot fish, Oplegnathus fasciatus (돌돔 (Oplegnathus fasciatus) 아가미의 미세구조)

  • Kim, Jae-Won;Baeck, Gun-Wook
    • Applied Microscopy
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    • v.34 no.2
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    • pp.139-143
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    • 2004
  • The ultrastructure of the gills of Oplegnathus fasciatus was examined by means of light and transmission electron microscopes. The gills have primary and secondary filaments (lamellae). The following cells are identified and described : epithelial cell, pillar cell, chloride cell and mucose cell. The simple epithelial layer consists of squamous epithelium containing a large nucleus and the surface is covered with some of microridges. The lamella pillar structures are characterized by axial microtubules and lateral membrane interdigitations. Chloride cells contain a lot of mitochondria and specifically developed tubular systems. The rough endoplasimic reticulum and golgi complex, and some of mucous granules were observed in immature mucous cells. The mature mucous cells were AB-PAS positive, globular in shape, and had mucous granules of similar size with various electron densities.

Efficient method for generating homozygous embryonic stem cells in mice

  • Kim, Bitnara;So, Seongjun;Choi, Jiwan;Kang, Eunju;Lee, Yeonmi
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.1
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    • pp.48-54
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    • 2022
  • Parthenogenesis is maternally uniparental reproduction through the embryonic development of oocytes without fertilization. Artificial activation of mature oocytes could generate homozygous haploid embryos with the extrusion of the second polar body. However, the haploid embryos showed low embryo development in preimplantation embryos. In this study, we investigated whether the electronic fusion of the haploid embryos could enhance embryo development and ESC establishment in mice. Haploid embryos showed the developmental delay from 4-cell to the blastocyst stage. The haploid blastomeres of the 2-cell stage were fused electronically, resulting in that the fused embryos showed a significantly higher rate of blastocysts compared to non-fused haploid embryos (55% vs. 37%). Further, the embryonic stem cells (ESCs) derived from the fused embryos were confirmed to be diploid. The rate of ESC establishment in fused embryos was significantly higher compared to non-fused ones. Based on the results, we concluded that the electronic fusion of haploid embryos could be efficient to generate homozygous ESCs.