• 생명과학회지
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• 제10권2호
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• pp.125-130
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• 2000

To overproduce endoxylanase from a recombinant Bacillus subtilis harboring the pJHKJ4 plasmid, the effects of carbon and nitrogen sources on the cell growth and expression level of endoxylanase were investigated in the flask cultures. Among the various carbon and nitrogen sources tested, glucose and maltose as carbon source and yeast extract as nitrogen source were found to be the most effective for the cell growth and the endoxylanase expression. When the concentration of glucose was increased from 0.5% to 5%, the highest activity of extracellular endoxylanse, 166 unit/$m\ell$, was observed at 2% glucose. In case of maltose, the endoxylanase was stably produced at the level of 180 unit/$m\ell$, regardless of the concentration of maltose. The higher the concentration of yeast extract, the greater cell growth and endoxylanase expression were obtained. However, the highest endoxylanase activity per unit cell mass was observed with 1% yeast extract. With the optimized medium (2% glucose, 1% yeast extract, etc), about 630 unit/$m\ell$ of endoxylanse was expressed through the batch fermentation in a fermentor, which expression level corresponded to about 0.7 g-endoxylanase protein /$\ell$. It was also found that the plasmid was stably maintained above 70% level, and more than 90% of endoxylanase activity was detected in the extracellular medium.

• Molecules and Cells
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• 제45권9호
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• pp.610-619
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• 2022

Cellular senescence plays a paradoxical role in tumorigenesis through the expression of diverse senescence-associated (SA) secretory phenotypes (SASPs). The heterogeneity of SA gene expression in cancer cells not only promotes cancer stemness but also protects these cells from chemotherapy. Despite the potential correlation between cancer and SA biomarkers, many transcriptional changes across distinct cell populations remain largely unknown. During the past decade, single-cell RNA sequencing (scRNA-seq) technologies have emerged as powerful experimental and analytical tools to dissect such diverse senescence-derived transcriptional changes. Here, we review the recent sequencing efforts that successfully characterized scRNA-seq data obtained from diverse cancer cells and elucidated the role of senescent cells in tumor malignancy. We further highlight the functional implications of SA genes expressed specifically in cancer and stromal cell populations in the tumor microenvironment. Translational research leveraging scRNA-seq profiling of SA genes will facilitate the identification of novel expression patterns underlying cancer susceptibility, providing new therapeutic opportunities in the era of precision medicine.

• Applied Microscopy
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• 제18권2호
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• pp.1-20
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• 1988

The species of the slug used in the experiment is Limax flavus L. For identifying the chemical characteristics of the epidermis, granules and mucus-producing cell of this animal is examined with methylene blue-basic fuchsin double stain and PAS-alcian blue reagent. For the ultrastructural research of the epidermal free surface, the epitheial cell and the parenchymal cell are used with scanning electron microscope and transmission elec-tron microscope respectively. I . Epidermal tissue The epidermal tissue of the slug is observed being divided into the dorsal and the ventral side(toot pad) respectively. 1) Dorsal epidermal tissue The dorsal epidermis of the slug is constituted with the simple columnar epithelium and the microvilli are compacted on the epidermal free surface. Two different types of the secretory granules of the neutral and the acid mucus are observed between the epithelial cells, and the neutral mucous granules are highest electron-dense but the acid mucous granules are observed to be electron-lucent. 2) Foot epidermal tissue The Foot epidermis is formed with the taller simple columnar epithelium than the dorsal epidermis and these cells have both a large number of the microvilli and a few number of the large villi. The secretory granules of three different types, which are acid, neutral and mixed mucous granule of two different types are observed between the epithelial cells. The neutral mucous granules are highest electron dense but the acid mucous granules are observed to be electron-lucent. II . Mucous granule-producing cell and mucus-producing cells Seven different types of the granules-producing cell and the mucus-producing cells are observed between the parenchyma. 1) A-type of acid mucous granule-producing cell The electron-lucent granules are largely occupied in the cytoplasm of these cells and then the granules are surrounded by irregular membrane. These electron-lucent granules exhibit alcianophilia with PAS-alcian blue reaction, so these granules are certified to be acid mucopolysaccharide. 2) B-type of acid mucus-producing cell The nucleus and the cytoplasm of these cells are pushed by the acid mucus of the electron-lucent toward the cell membrane. This mucus has been confirmed to be the acid mucopolysaccharide with PAS-alcian blue reagent. 3) A-type of neutral mucous granule-producing cell These cells contain the electron-dense round granules with approximately $1{\mu}m$ in diameter, which exhibit strongly PAS-positive reaction. These granules are confirmed to be the neutral mucoplysaccharide. 4) B-type of neutral mucous granule-producing cell These cells contain two different types of electron dense granules and electron-lucent granules; The former exhibits to be strongly PAS-positive and the latter to have alcianophilia reaction respectively. 5) C-type of neutral mucus-producing cell These cells are similar to the shape and the size of the B-type of mucus-producing cell but these two different types of cells are stained with reversing properties to each other. The mucus of the C-type cell that electron-lucent is largely occupied in the cytoplasm that exhibits strongly PAS-positive reaction. 6) D-type of neutral mucous granule-producing cell These cells contain round granules about $1{\mu}m$ in size which are observed to be medium electron-dense granules and those granules are stained brightly red with PAS-weak positive reaction. The granules are certified to be neutral mucopolysaccharide. 7) E-type of neutral mucous granule-producing cell These cells are similar to the shape and the size of the D-type of neutral mucous granule-producing cell. These cells contain a large number of granules with about $1{\mu}m$ in diameter showing electron-lucent and then granules are seen to be PAS-weak positive reaction. III. Parenchyma The clear cell and dark cell are found in the parenchyma of the Limax flavus L. 1) Clear cell These cells are round formed and the nucleus of the cells are larger than cytoplasm. These cells which have the electron-lucent cytosol possess poorly developed organelles. 2) Dark cell These cells are found to be dark cells due to high electron-density, which exhibit strongly methylene-blue reaction from double stain of methylene blue-basic fuchsin.

• Clinical and Experimental Reproductive Medicine
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• 제31권1호
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• pp.19-27
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• 2004

Objective: This study was to examine the in vitro neural cell differentiation patterns of human embryonic stem (hES) cells following treatment of various neurotrophic factors [basic fibroblast growth factor (bFGF), retinoic acid (RA), brain derived neurotrophic factor (BDNF) and transforming growth factor (TGF)-$\alpha$], particulary in dopaminergic neuron formation. Methods: The hES cells were induced to differentiate by bFGF and RA. Group I) In bFGF induction method, embryoid bodies (EBs, for 4 days) derived from hES were plated onto gelatin dish, selected for 8 days in ITSFn medium and expanded at the presence of bFGF (10 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14 and 21 days. Group II) For RA induction, EBs were exposed of RA ($10^{-6}M$) for 4 days and allowed to differentiate in N2 medium for 7, 14 and 21 days. Group III) To examine the effects of additional neurotrophic factors, bFGF or RA induced cells were exposed to either BDNF (10 ng/ml) or TGF-$\alpha$ (10 ng/ml) during the 21 days of final differentiation. Neuron differentiation and dopamine secretion were examined by indirect immunocytochemistry and HPLC, respectively. Results: The bFGF or RA treated hES cells were resulted in similar neural cell differentiation patterns at the terminal differentiation stage, specifically, 75% neurons and 11% glial cells. Additionally, treatment of hES cells with BDNF or TGF-$\alpha$ during the terminal differentiation stage led to significantly increased tyrosine hydroxylase (TH) expression of a dopaminergic neuron marker, compared to control (p<0.05). In contrast, no effect was observed on the rate of mature neuron (NF-200) or glutamic acid decarboxylase-positive neurons. Immunocytochemistry and HPLC analyses revealed the higher levels of TH expression (20.3%) and dopamine secretion (265.5 $\pm$ 62.8 pmol/mg) in bFGF and TGF-sequentially treated hES cells than those in $\alpha$ RA or BDNF treated hES cells. Conclusion: These results indicate that the generation of dopamine secretory neurons from in vitro differentiated hES cells can be improved by TGF-$\alpha$ addition in the bFGF induction protocol.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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• pp.1919-1927
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• 2006

Tumor cells genetically engineered to secrete cytokines are effective in tumor therapy, but various unexpected side effects are observed, which may result from the bulk activation of various bystander cells. In this study, we tested tumor vaccines expressing various membrane-bound forms of IL-2 (mbIL-2) on MethA fibrosarcoma cells to focus antitumor immune responses to CTL. Chimeric forms of IL-2 with whole CD4, deletion forms of CD4, and TNF were expressed on the tumor cell surface, respectively. Tumor clones expressing mbIL-2 or secretory form of IL-2 were able to support the cell growth of CTLL-2, an IL-2-dependent T cell line, and the proliferation of spleen cells from 2C TCR transgenic mice that are responsive to the $p2Ca/L^d$ MHC class I complex. Expression of mbIL-2 on tumor cells reduced the tumorigenicity of tumor cells, and the mice that once rejected the live IL-2/TNF tumor clone acquired systemic immunity against wild-type MethA cells. The IL-2/TNF clone was inferior to other clones in tumor formation, and superior in the stimulation of the CD8+ T cell population in vitro. These results suggest that the IL-2/TNF clone is the best tumor vaccine, and may stimulate CD8+ T cells by direct priming. Expression of IL-2/TNF on tumor cells may serve as an effective gene therapy method to ameliorate the side effects encountered in the recombinant cytokine therapy and the conventional cytokine gene therapy using the secretory form of IL-2.

• Applied Microscopy
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• 제35권2호
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• pp.97-104
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• 2005

표피조직은 비교적 분화되지 않은 기본 표피세포와 식물에 따라 특수하게 분화되어 형태와 기능이 매우 다른 여러 이형세포로 발달할 수 있다. 본 연구에서는 약용으로 활용되는 접시꽃 (Althaea rosea)의 엽육 표피조직에 발달하는 모용에 대하여 이들 모용 유형에 따른 분화 발달 양상을 주사전자현미경적으로 연구하였다. 접시꽃 엽육 표피상에 발달하는 모용은 크게 단모(simple hairs), 총생모 (non-grandular tufted hairs)와 분비모 (peltate glandular hairs)로 대별되는데, 총생모는 상, 하피 및 하피의 엽맥을 따라 특히 더 발달하는 장상의 총생모 (longtufted hairs, ${\sim}1000{\mu}m,\;2{\sim}10$ branchlet)와 상, 하피 및 하피의 엽신에 더 밀생 분포하는 단상의 총생모 (shorttufted hairs, ${\sim}210{\mu}m,\;2{\sim}7$ branchlet)로 나뉘어진다. 장상의 총생모는 단순모로부터 시작하여 10개까지 분지되며, 분화 초기에는 비교적 일정한 방향으로 분지되는 양상을 보인다. 반면 분비모는 $1{\sim}2$개의 두정세포(head cell, 직경 $20{\sim}35{\mu}m$), 2개의 자루세포(stalk cell, 길이 $10{\sim}15{\mu}m$), 기저세포 (basal cell)로 이루어진다. 또한, 엽신 전체에 발달하는 총생모와는 달리 분비모는 특히 표피면에서 함몰된 망목극(areole)을 따라 분포한다. 이들 모용 중 총생모는 엽육 표피의 보호 및 방어기능을 주로 수행할 것으로 추측되며, 분비모는 두정세포 내에 함유되어 있는 성분이 유용한 이차대사 물질을 분비하는 기능과 보호기능에도 중요한 역할을 하는 것으로 추정된다. 일반적으로 모용은 분비기능이 활발한 식물에서 유용한 이차대사 화합물을 생성, 축적하고 분비하는데 중추적인 기능을 수행하는 것으로 알려져 있어 이들에 대한 세포수준에서의 연구 및 분비되는 유용한 물질에 대한 연구는 구조적 정보 및 적응을 위한 방어기작 규명 등에 이르기까지 매우 유용하게 쓰일 것이다.

• 한국동물학회지
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• 제33권1호
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• pp.22-27
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• 1990

본 연구는 옴개구리의 위저부 점막을 전자현미경으로 관찰한 결과, 내분비세포의 분비 과립의 크기, 형태, 전자밀도 및 세포의 형태에 따라서 다음과 같이 3종류의 세포를 분류하였다. 제 I 형 : 전자밀도가 높고 원형 또는 난원혀으이 과립을 가지며, 과립내용물과 한계막 사이의 편제하거나 공포상으로 출현하였다. 과립의 크기는 직경이 300-500nm였다. 제 II 형 : 구형 또는 난원형의 과립을 가지며, 전자밀도는 낮거나 높게 나타나며, 한계막과 과립내용물 사이에는 좁은 halo를 나타내었다. 과립의 크리는 직경이 110-230nm였다. 제 III 형 : 신장된 난원형 또는 다형태성의 과립을 가지며, 전자밀도는 낮거나 중등도이며 과림가 내용물 사이에는 명확한 halo를 형성하고 세포질내에는 미세섬유들이 풍부하게 출현하였다. 과립의 크기는 직경이 50-200nm였다.

• Biomolecules & Therapeutics
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• 제29권3호
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• pp.342-351
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• 2021

Liver colonization is initiated through the interplay between tumor cells and adhesion molecules present in liver sinusoidal endothelial cells (LSECs). This crosstalk stimulates tumor COX-2 upregulation and PGE2 secretion. To elucidate the role of the LSEC intercellular adhesion molecule-1 (ICAM-1) in the prometastatic response exerted by tumor and stromal COX-2, we utilized celecoxib (CLX) as a COX-2 inhibitory agent. We analyzed the in vitro proliferative and secretory responses of murine C26 colorectal cancer (CRC) cells to soluble ICAM-1 (sICAM-1), cultured alone or with LSECs, and their effect on LSEC and hepatic stellate cell (HSC) migration and in vivo liver metastasis. CLX reduced sICAM-1-stimulated COX-2 activation and PGE2 secretion in C26 cells cultured alone or cocultured with LSECs. Moreover, CLX abrogated sICAM-1-induced C26 cell proliferation and C26 secretion of promigratory factors for LSECs and HSCs. Interestingly, CLX reduced the protumoral response of HSC, reducing their migratory potential when stimulated with C26 secretomes and impairing their secretion of chemotactic factors for LSECs and C26 cells and proliferative factors for C26 cells. In vivo, CLX abrogated the prometastatic ability of sICAM-1-activated C26 cells while reducing liver metastasis. COX-2 inhibition blocked the creation of a favorable tumor microenvironment (TME) by hindering the intratumoral recruitment of activated HSCs and macrophages in addition to the accumulation of fibrillar collagen. These results point to COX-2 being a key modulator of processes initiated by host ICAM-1 during tumor cell/LSEC/HSC crosstalk, leading to the creation of a prometastatic TME in the liver.

• 한국동물학회지
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• 제16권1호
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• pp.13-23
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• 1973

저자들은 성숙한 雄性白鼠(Albino Wistar)에 INH 40mg/kg, pyridoxine 20mg/kg를 경구적으로 투여하여 mast cell과 enterochromaffin cell 의 변동을 관찰하여 다음과 같은 결과를 얻었다. 1. INH투여로 白鼠 약 舌 mast cell 의 심한 파괴와 細胞質顆粒의 逸出作用으로 세포수는 격감하였으나, pyridoxine 투여군에서는 대조군에 비해 상당한 수적증가를 보였다. 2. INH 투여로 白鼠 심이지장 enterochromaffin cell의 세포수와 細胞質顆粒양에 현저한 감소를 나타냈으나 pyridoxine 투여군에서는 대조군에 비해 enterochromaffin cell의 상당한 수적증가를 보였다. 3. 이상의 성적에 있어서 INH의 다량 투여는 mast cell과 enterochromaffin cell의 생성에 심한 장해를 주며 pyridoxine 의 적당한 투여는 mast cell과 enterochromaffin cell의 생성을 조장 하고 顆粒양을 증가시키는 것으로 보아 이들 세포의 分泌産物인 5-Hydroxytryptamine 대사에 pyridoxine이 관여하는 것으로 사료된다.

• BMB Reports
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• 제53권2호
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• pp.65-73
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• 2020

Life expectancy has dramatically increased around the world over the last few decades, and staying healthier longer, without chronic disease, has become an important issue. Although understanding aging is a grand challenge, our understanding of the mechanisms underlying the degeneration of cell and tissue functions with age and its contribution to chronic disease has greatly advanced during the past decade. As our immune system alters with aging, abnormal activation of immune cells leads to imbalance of innate and adaptive immunity and develops a persistent and mild systemic inflammation, inflammaging. With their unique therapeutic properties, such as immunomodulation and tissue regeneration, mesenchymal stem cells (MSCs) have been considered to be a promising source for treating autoimmune disease or as anti-aging therapy. Although direct evidence of the role of MSCs in inflammaging has not been thoroughly studied, features reported in senescent MSCs or the aging process of MSCs are associated with inflammaging; MSC niche-driven skewing of hematopoiesis toward the myeloid lineage or oncogenesis, production of pro-inflammatory cytokines, and weakening their modulative property on macrophage polarization, which plays a central role on inflammaging development. This review explores the role of senescent MSCs as an important regulator for onset and progression of inflammaging and as an effective target for anti-aging strategies.