• Title/Summary/Keyword: Primary culture

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In Vitro Culture of Human Nasal Epithelial Cells by Monolayer Culture of Dissociated Cells (분리 세포의 단층세포 배양법에 의한 인체 비점막 상피세포의 배양)

  • Kim, Yong-Dae;Song, Si-Youn;Min, Myung-Ki;Sub, Jang-Su;Song, Kei-Won;Park, Ho-Sun
    • Journal of Yeungnam Medical Science
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    • v.15 no.2
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    • pp.286-296
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    • 1998
  • Different techniques for culturing respiratory epithelial cells have been developed to overcome the limitations of studies on in vivo and on bioptic material. Traditionally, culture systems are divided into organ cultures, explant cultures and dissociated cell cultures. The first two contain both epithelial and non-epithelial cells. However, in monolayer cultures of dissociated cells only epithelial cells are present, the effects observed are caused by a pure epithelial responses. The purpose of this study is to establish primary culture method of human nasal epithelium (HNEC) by monolayer culture of dissociated cells to evaluate the role of the epithelial cells in the allergic and non-allergic nasal inflammatory reactions. HNEC was prepared by primary culture method of monolayer culture of dissociated cells from human inferior nasal turbinate mucosa of septal deviation patients. Primary cultured cells were characterized by indirect immunofluorescence assay and transmission electron microscopy. The immunoreactivities of cytokeratin-pan and cytokeratin No. 8 were observed in cultured HNEC. However, the immnoreactivities of vimentin and von Willebrand factor were not observed in cultured HNEC. The tonofilaments and desmosome were observed in cultured HNEC. The cultured epithelial cells were identified to be pure nasal epithelial cells. The monolayer culture of dissociated cells could successfully be employed for further study to investigate the role of the epithelial cells in allergic or non-allergic nasal inflammatory diseases.

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Effects of the Methanol Extract of Bupleuri Radix on Primary Cultured Brain Cells, DRG and Hepatocytes (시호의 메탄올 추출물이 일차배양한 뇌, DRG 및 간세포에 미치는 영향)

  • Kim, Young-Choong;Park, Mi-Jung;Byun, Soon-Jung;Song, Jin-Ho
    • Korean Journal of Pharmacognosy
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    • v.21 no.1
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    • pp.92-99
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    • 1990
  • Effects of the methanol extract of Bupleuri Radix on primary cultured chicken embryonic brain cells, dorsal root ganglia (DRG) and rat hepatocytes were studied. The methanol extract of Bupleuri Radix at the concentration ranging from $10{\;}{\mu}g/ml\;to\;100{\;}{\mu}g/ml$ significantly recovered the cytotoxicity of rat hepatocytes induced by the treatment of galactosamine; at the concentration of $100\;{\mu}g/ml$, values of GOT and GPT in the culture medium were reduced by the 60% and 75%, respectively of those in the absence of the methanol extract of Bupleuri Radix. The addition of the methanol extract of Bupleuri Radix. into chicken embryonic brain cells which were cultured with a deficient medium significantly increased the number of cells promoting the neurite outgrowth. However, the methanol extract of Bupleuri Radix showed no effect on the activities of PDHC and acetylcholinesterase in primary cultured brain cells.

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The Influence of Culture on the Experiences of Korean, Korean American, and Caucasian-American Family Caregivers of Frail Older Adults: A Literature Review

  • Kong, Eun-Hi
    • Journal of Korean Academy of Nursing
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    • v.37 no.2
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    • pp.213-220
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    • 2007
  • Purpose. The purpose of this review is to explore cultural influences on the experiences of Korean, Korean American, and Caucasian American family caregivers caring for frail older adults in terms of the selection of a primary caregiver, caregiving motivation, support/help-seeking, and negative emotional responses (depression and burden). Methods. Seven electronic databases were searched to retrieve studies from 1966 to 2005. Thirty-two studies were identified. Results. This review supported cultural influences on the selection of primary caregiver, caregiving motivation, and support/help-seeking among the three caregiver groups. In Korean caregivers, the major primary caregivers were daughters-in-law while among Korean American and Caucasian American caregivers, the major primary caregivers were daughters or spouses. As a major caregiving motivation, Caucasian American care¬givers reported filial affection while Korean caregivers and Korean American caregivers reported filial obligation. Korean caregivers reported higher extended family support, while Caucasian American caregivers reported higher utilization of formal support. Korean caregivers showed the highest levels of depression followed by Korean American caregivers and Caucasian American caregivers. Conclusion. In order to develop culturally appropriate interventions and policies, more research is needed to further explain these differences among the three groups, especially regarding support/help-seeking and negative emotional responses.

Induction of Differentiation of the Cultured Rat Mammary Epithelial Cells by Triterpene Acids

  • Paik, Kee-Joo;Jeon, Seong-Sill;Chung, Hae-Young;Lee, Kyung-Hee;Kim, Kyu-Won;Chung, Joon-Ki;Kim, Nam-Deuk
    • Archives of Pharmacal Research
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    • v.21 no.4
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    • pp.398-405
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    • 1998
  • We investigated the effects of triterpene acids (TAs), ursolic acid (UA) and oleanolic acid (OA), on the induction of proliferation and differentiation of normal rat mammary epithelial cells (RMEC) or organoids cultured in Matrigel or primary culture system. To elucidate the effects, we tested their differentiation inducing activities with intercellular communication ability, cell cycle patterns, induction of apoptosis, and morphological differentiation in the three dimensional extracellular culture system. To study the changes of RMEC subpopulation in culture, the cultured cells were isolated, immunostained with peanut lectin (PNA) and anti-Thy-1.1 antibody and then analyzed with flow cytometry. Four different subpopulations, such as PNA and Thy-1.1 negative cells (B-), PNA positive cells (PNA+), Thy-1.1 positive cells (Thy-1.1+), PNA and Thy-1.1 positive cells (B+), were obtained and the size of each subpopulation was changed in culture with time in the presence of TAs. Intercellular communication was observed in culture for 7 days in TAs-treated cells, but not in culture for 4 days with scrape-loading dye transfer technique. $G_2$/M phase cells and the number of apoptotic population were increased in TAs-treated groups in cell cycle analyses. S phase fractions were reduced and the change of $G_1$ phase cells was not observed. The colonies with distinct multicelfular structures, such as stellate, ductal, webbed, squamous, lobulo-ductal colonies, were observed in Matrigel culture and the frequencies of each colony were changed in the presence of TAs. These results suggest that UA and OA have differentiation inducing effects on rat mammary epithelial cells in primary or in Matrigel culture.

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Recombinant Human Parathyroid Hormone Related Peptide (1-34) Stimulates Osteoclastic Bone Resorption in Both Rodent and Avian Disagsresated Osteoclast Culture (파골세포배야에서 나타난 부갑상선호르몬의 설치류 및 조류 파골세포에 대한 촉진 효과)

  • 양대석;김일찬남궁용이창호
    • The Korean Journal of Zoology
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    • v.37 no.2
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    • pp.255-261
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    • 1994
  • Recombinant human pBrathyriod hormone related peptide (1-341 (rhPTHrP) has been known to stimulate bone resorption in intact bone tissue culture system. Osteoclast has been known as a primary responsible cell for bone resorption. To examine the effect of rhPTHrP on this cell, we employed disaggregated rat osteodast culture. As a result, we found that rhPTHrP sisnificBntly elevates both the number and total area of resorbed pits in this culture. On the other hand, the conflicting results between disagsregated rat osteoc13st culture and Ca2+-deficient hen osteoclast culture system have been a big obstacle for the progress of bone research. To verify the differences between rat 3nd chick osteoclast system, we performed the same experiment using chick embryonic osteoclast. Since the similar results were obtained from the disaggregated chick osteoclast culture, the discrepancy between chick and rat osteoclast culture study seemed to be due to the difference in culture method, rather due to the species-difference.

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Studies on the Air-Liquid Interface Culture as an Experimental Model for Physiology and Pharmacology of Tracheal Epithelial Cells (기관(氣管) 상피세포 생리 및 약리 실험모델로서의 공기-액체 접면 일차배양법 연구)

  • 이충재;이재흔;석정호;허강민
    • Biomolecules & Therapeutics
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    • v.10 no.4
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    • pp.281-286
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    • 2002
  • In this study, we intended to get a preliminary data for establishing rat tracheal surface epithelial(RTSE) cell culture system as an experimental model for physiology and pharmacology of tracheal epithelial cells. Primary culture on the membrane support and application of the air-liquid interface system at the level of cell layer were performed. The cell growth rate and mucin production rate were measured according to the days in culture. The results were as follows: this culture system was found to manifest mucocilliary differentiation of rat tracheal epithelial cells, the cells were confluent and the quantity of produced and released mucin was highest on culture day 9, the mucin was mainly released to the apical side and tbe free $^3{H}$-glucosamine which was not incorporated to process of synthesis of mucin was left on the basolateral side. Taken together, we suggest that air-liquid interface culture system can be used as a substitute for immersion culture system and as an experimental model for in vivo mucus-hypersecretory diseases.

Development of a Primary Tissue Culture Method having Greater Reliability than Isolated Cell Cultures - Steroid-Responsiveness of Uterine Myometrial and Myomatous(Leiomyomatous) Cells (자궁근종세포의 최적 초기배양 조건 확립 - 정상 자궁근세포와 자궁근종세포의 스테로이드에 대한 반응)

  • Lee, Eun-Ju;Bajracharya, Prati;Hyun, Jin-Hee;Kim, Hang-Jin;Song, Gun-Ho;Cho, Kyung-Hyun;Lee, Dong-Mok;Lee, Taek-Hoo;Chun, Sang-Sik;Choi, In-Ho
    • Development and Reproduction
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    • v.11 no.3
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    • pp.205-217
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    • 2007
  • As an initial step toward better understanding of the molecular mechanism of estrogen-dependent growth in myoma, an optimal primary cell culture condition has been developed and examined by this study. Myoma and myometrium were cultured by two different methods. Culture stability and $E_2$-responsiveness in stable culture were studied. The culture of digested tissue pieces(Method 2) was found to be a stable culture method for the myoma and myometrium showing a favorable response to estrogen. mRNA expression of PR, IGF-1 and IGF-1 receptor genes was enhanced by $E_2$. The gene responses to $E_2$ were higher in myoma compared with myometrium. Moreover, these responses were more expressive in tissues than in the surrounding cells in primary culture of normal myometrium and myoma, implying a vital role of cell communication through the extracellular matrix in maintaining the estrogen-responsiveness. The development of an improved cell culture system for myoma provides an in vitro tool to further investigate the basis of the tumor formation.

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Development of Isolation and Cultivation Method for Outer Root Sheath Cells from Human Hair Follicle and Construction of Bioartificial Skin

  • Sin, Yeon-Ho;Seo, Yeong-Gwon;Lee, Du-Hun;Yu, Bo-Yeong;Song, Gye-Yong;Seo, Seong-Jun;Hwang, Seong-Ju;Kim, Yeong-Jin;Yang, Eun-Gyeong;Park, Jang-Seo;Jang, Lee-Seop;Park, Jeong-Geuk
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.302-305
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    • 2003
  • It is difficult to obtain sufficient healthy skin for coverage of a wide area of skin wound. In the skin, an additional population of living epithelial cells is located in the outer root sheath (ORS) of hair $follicles.^{1),2)}$ ORS cells should be a good source of epithelium because they are easily obtainable and patients do not have to suffer from scar formation at donor sites. We modified ordinary primary culture technique for the purpose of solving such problem that epithelial cells have a low propagation and easy aging during culture periods. First of all, we improved primary cultivation methods. In the ordinary primary culture, average yield of human ORS cells was $2\;{\times}\;10^3$ cells/follicle by direct incubation with trypsin (0.1%)/EDTA (0.02%) solution for 15 min at $37^{\circ}C$ but we could obtain about $6.5\;{\times}\;10^3$ cells/follicle by two step enzyme digestion method with dispase (1.2 U/ml) and trypsin (0.1%)/EDTA (0.02%) solution. So we could achieve three times higher primary cultured ORS cell yield. Secondly, we could obtain total $2\;{\times}\;10^7$ cells in serum free medium and even more total $6\;{\times}\;10^7$ cells in modified E-medium with mitomycin C-treated feeder cells during 17 days. Using the cultured ORS cells, and we could make bioartificial skin equivalent in vitro and concluded that ORS cells were progenitor cells for skin epithelial cell.

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Factors Affecting Primary Culture of Nuclear Transfer Blastocysts for Isolation of Embryonic Stem Cells in Miniature Pigs

  • Kim, Min-Jeong;Ahn, Kwang-Sung;Kim, Young-June;Shim, Ho-Sup
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.133-137
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    • 2009
  • Pluripotent embryonic stem (ES) cells isolated from inner cell mass (ICM) of blastocyst-stage embryos are capable of differentiating into various cell lineages and demonstrate germ-line transmission in experimentally produced chimeras. These cells have a great potential as tools for transgenic animal production, screening of newly-developed drugs, and cell therapy. Miniature pigs, selectively bred pigs for small size, offer several advantages over large breed pigs in biomedical research including human disease model and xenotransplantation. In the present study, factors affecting primary culture of somatic cell nuclear transfer blastocysts from miniature pigs for isolation of ES cells were investigated. Formation of primary colonies occurred only on STO cells in human ES medium. In contrast, no ICM outgrowth was observed on mouse embryonic fibroblasts (MEF) in porcine ES medium. Plating intact blastocysts and isolated ICM resulted in comparable attachment on feeder layer and primary colony formation. After subculture of ES-like colonies, two putative ES cell lines were isolated. Colonies of putative ES cells morphologically resembled murine ES cells. These cells were maintained in culture up to three passages, but lost by spontaneous differentiation. The present study demonstrates factors involved in the early stage of nuclear transfer ES cell isolation in miniature pigs. However, long-term maintenance and characterization of nuclear transfer ES cells in miniature pigs are remained to be done in further studies.