• Title/Summary/Keyword: primary culture

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Insulin enhances neurite extension and myelination of diabetic neuropathy neurons

  • Pham, Vuong M.;Thakor, Nitish
    • The Korean Journal of Pain
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    • v.35 no.2
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    • pp.160-172
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    • 2022
  • Background: The authors established an in vitro model of diabetic neuropathy based on the culture system of primary neurons and Schwann cells (SCs) to mimic similar symptoms observed in in vivo models of this complication, such as impaired neurite extension and impaired myelination. The model was then utilized to investigate the effects of insulin on enhancing neurite extension and myelination of diabetic neurons. Methods: SCs and primary neurons were cultured under conditions mimicking hyperglycemia prepared by adding glucose to the basal culture medium. In a single culture, the proliferation and maturation of SCs and the neurite extension of neurons were evaluated. In a co-culture, the percentage of myelination of diabetic neurons was investigated. Insulin at different concentrations was supplemented to culture media to examine its effects on neurite extension and myelination. Results: The cells showed similar symptoms observed in in vivo models of this complication. In a single culture, hyperglycemia attenuated the proliferation and maturation of SCs, induced apoptosis, and impaired neurite extension of both sensory and motor neurons. In a co-culture of SCs and neurons, the percentage of myelinated neurites in the hyperglycemia-treated group was significantly lower than that in the control group. This impaired neurite extension and myelination was reversed by the introduction of insulin to the hyperglycemic culture media. Conclusions: Insulin may be a potential candidate for improving diabetic neuropathy. Insulin can function as a neurotrophic factor to support both neurons and SCs. Further research is needed to discover the potential of insulin in improving diabetic neuropathy.

Analysis of Needs for Sexual Education in Primary School Children (초등학교 고학년 학생의 성교육 요구 내용 분석)

  • Yang Soon Ok;Jeong Geum Hee;Han Young Ran
    • Child Health Nursing Research
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    • v.7 no.1
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    • pp.5-20
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    • 2001
  • The purpose of this study was to analyze and identify the higher-grade primary school children's needs for sexual education. We got the data with open question from 481 children from March to July 1999. We analyzed them according to the framework suggested by the Sex Information and Education Council of USA. The framework involves the six domains such as human development, relationship, personal skills, sexual behavior, sexual health, and society & culture. In the analysis of need for sexual education according to domain, need for human development was highest (79.2 %), after that followed society & culture (8.3 %), sexual health (5.8%), sexual behavior (3.6%), and relationships (3.1%). There was no need for personal skills. By topics, there were needs for puberty (38.0%), reproduction (32.5%), gender roles (6.0%), body image (4.9%), sexual abuse (2.4%) and reproductive health (2.4%). Girls and boys all mostly wanted to know human development 41.0 % and 38.2 % each. Society & culture (3.4 %), sexual behaviour (2.7%), sexual health (2.6%), relationship (2.4%) were domains selected by boys. Girls wanted to know society & culture (4.9%) and 'sexual health (3.2%). Comparing by grade, fourth, fifth and sixth grade children are commonly interested in development, society & culture. Fourth grade children scarcely show attention on the relationship, sexual behaviour and sexual health; meanwhile sixth grade children would like to know sexual behaviour and sexual health. When their grade became higher, they would like to know more precise, concrete and deep information. We suggest that the textbook/guidebook for sexual health for the higher-grade primary school children should be developed considering the sex and grade.

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Primary Culture of Human Nasal Epithelial Cell Monolayer for In Vitro Drug Transport Studies (약물의 in vitro 투과 실험을 위한 사람의 비강점막상피세포 단층막의 일차배양)

  • Yoo, Jin-Wook;Kim, Yoo-Sun;Lee, Min-Ki;Roh, Hwan-Jung;Lee, Chi-Ho;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.32 no.1
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    • pp.21-26
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    • 2002
  • The primary culture of human nasal epithelial cell monolayer was performed on a Transwell. The effect of various factors on the tight junction formation was observed in order to develop an in vitro experimental system for nasal transport studies. Human nasal epithelial cells, collected from human normal inferior turbinates, were plated onto diverse inserts. After 4 days, media of the apical surface was removed for air-liquid interface (ALI) culture. Morphological characteristics was observed by transmission electron microscopy (TEM). A polyester membrane of $0.4\;{\mu}m$ pore size was determined as the most effective insert based on the change in the transepithelial electric resistance (TEER) value as well as the $^{14}C-mannitol$ transport study. The ALI method was effective in developing the tight junction as observed in the further increase in the TEER value and reduction in the permeability coefficient $(P_{app})$ of $^{14}C-mannitol$ transport. Results of the transport study of a model drug, budesonide, showed that the primary culture system developed in this study could be further developed and applied for in vitro nasal transport studies.

A Consideration on the culture of mathematics classrooms (수학 교실 문화에 관한 소고)

  • 김남균
    • Education of Primary School Mathematics
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    • v.5 no.2
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    • pp.163-172
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    • 2001
  • Recently, higher discussions are made up on the culture of mathematics classrooms. These discussions are mostly about desirable culture and reform of culture in mathematics education. This paper is to offer a view point of seeing the culture of mathematics classroom. We need to look straightly our classroom as well as to reform the culture of mathematice classroom. This paper recommend sociomathematical norms to analyze and reform classroom culture. A elementary mathematics classroom was analyzed to understand our own culture of mathematics classroom.

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Immortalization of Primary Keratinocytes and Its Application to Skin Research

  • Choi, Moonju;Lee, Choongho
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.391-399
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    • 2015
  • As a major component of the epidermal tissue, a primary keratinocyte has served as an essential tool not only for the study of pathogenesis of skin-related diseases but also for the assessment of potential toxicities of various chemicals used in cosmetics. However, its short lifespan in ex vivo setting has been a great hurdle for many practical applications. Therefore, a number of immortalization attempts have been made with success to overcome this limitation. In order to understand the immortalization process of a primary keratinocyte, several key biological phenomena governing its lifespan will be reviewed first. Then, various immortalization methods for the establishment of stable keratinocyte cell lines will be explained. Finally, its application to a three-dimensional skin culture system will be described.

Factors Affecting Primary Cultures of Abalone Haliotis discus hannai Ovary-dissociated Cells and General Culture Aspects

  • Ryu, Jun Hyung;Nam, Yoon Kwon;Gong, Seung Pyo
    • Fisheries and Aquatic Sciences
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    • v.18 no.1
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    • pp.81-88
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    • 2015
  • We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ${\geq}10$ weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture-18.4 and 19.5 days for first and second subcultures, respectively-and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

Isolation and In Vitro Culture of Vascular Endothelial Cells from Mice

  • Choi, Shinkyu;Kim, Ji Aee;Kim, Kwan Chang;Suh, Suk Hyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.1
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    • pp.35-42
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    • 2015
  • In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.

Effect of Glial-neuronal Cell Co-culture on GFAP Expression of Astrocytes (신경세포가 별아교세포의 아교섬유성 산단백질 표현에 미치는 영향)

  • Bae Hyung-Mi;Park Jung-Sun;Yeon Dong-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.3
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    • pp.285-296
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    • 1997
  • Injury to brain transforms resting astrocytes to their reactive form, the hallmark of which is an increase in glial fibrillary acidic protein (GFAP), the major intermediate filament protein of their cell type. The overall glial response after brain injury is referred to as reactive gliosis. Glial-neuronal interaction is important for neuronal migration, neurite outgrowth and axonal guidance during ontogenic development. Although much attention has been given to glial regulation of neuronal development and regeneration, evidences also suggest a neuronal influence on glial cell differentiation, maturation and function. The aim of the present study was to analyze the effects of glial-hippocampal neuronal co-culture on GFAP expression in the co-cultured astrocytes. The following antibodies were used for double immunostaining chemistry; mouse monoclonal antibodies for confirm neuronal cells, rabbit anti GFAP antibodies for confirm astrocytes. Primary cultured astrocytes showed the typical flat polygonal morphology in culture and expressed strong GFAP and vimentin. Co-cultured hippocampal neurons on astrocytes had phase bright cell body and well branched neurites. About half of co-cultured astrocytes expressed negative or weak GFAP and vimentin. After 2 hour glutamate (0.5 mM) exposure of glial-neuronal co-culture, neuronal cells lost their neurites and most of astrocytes expressed strong CFAE and vimentin. In Western blot analysis, total GFAP and vimentin contents in co-cultured astrocytes were lower than those of primary cultured astrocytes. After glutamate exposure of glial-neuronal co-culture, GFAP and vimentin contents in astrocytes were increased to the level of primary cultured astrocytes. These results suggest that neuronal cell decrease GFAP expression in co-cultured astrocytes and hippocampal neuronal-glial co-culture can be used as a reactive gliosis model in vitro for studying GFAP expression of astrocytes.

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In vitro Nasal Cell Culture Systems for Drug Transport Studies

  • Cho, Hyun-Jong;Termsarasab, Ubonvan;Kim, Jung-Sun;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.40 no.6
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    • pp.321-332
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    • 2010
  • Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the "passage" culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.

A Competitive Advantage Strategy Based on Innovative Culture and Quality of Work Life: Evidence from SMEs of the Tourism Industry in Indonesia

  • HERMAWATI, Adya;ANAM, Choirul;SUWARTA, Suwarta;WARDHANI, Arie Restu
    • The Journal of Asian Finance, Economics and Business
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    • v.9 no.8
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    • pp.29-36
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    • 2022
  • The objective of this research is to find out the effect of innovative culture and quality of work life on competitive advantage strategy with the mediation of individual performance. This research is the continuance of previous research conducted by Adya Hermawati with an originality aspect emphasizing a concept comprising innovative culture, quality of work life, and individual performance as factors that control competitive advantage strategy. The research subject is Tourism Industry SMEs. Explanatory research is a research method used in this study, by surveying respondents. The data sources in this research are primary and secondary. Primary data is collected from respondents directly through a questionnaire whereas secondary data are obtained from references that are relevant to research problems. In conformity with this explanation, the type of research data is quantitative data. The results of this research show that: innovative culture has an effect on individual performance, quality of work life affects individual performance, innovative culture has an effect on competitive advantage, quality of work life affects competitive advantage, individual performance has an effect on competitive advantage, innovative culture affects competitive advantage with the mediation of individual performance, and quality of work life affects competitive advantage with the mediation of individual performance.