• 제목/요약/키워드: STRO-1

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유세포 분석을 통한 과잉치 치수 유래 세포의 줄기세포 특성 연구 (Characterization of Human Dental Pulp Cells from Supernumerary Teeth by Using Flow Cytometry Analysis)

  • 류연숙;김종빈;신지선;이준행;김종수
    • 대한소아치과학회지
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    • 제46권3호
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    • pp.337-342
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    • 2019
  • 이 연구의 목적은 발치한 매복 과잉치에서 얻은 치수 유래 세포의 줄기세포 특성을 유세포 분석을 통해 알아보는 것이다. 15명의 환자로부터 채취한 정중 과잉치의 치수 세포를 계대 배양하였고, 3계대와 10계대 세포들을 유세포 분석을 이용하여 분석하였다. 간엽성 줄기세포 표지자 관찰에 사용된 항체 표지자(CD)는 양성 표지자로는 CD 73, CD 90, 그리고STRO-1 와 음성 표지자로는 CD 34, CD 45 이었다. 3계대에서 CD 73, CD 90은 각각 94.82%, 98.86%의 양성반응을, CD 34, CD 45는 각각 2.25%, 2.52%로 음성 반응을 보였으며, STRO-1은 20.93%를 나타냈다. 10계대에서는 CD 73, CD 90은 각각 96.62%, 98.61%의 발현을 보였지만, CD 34, CD 45는 각각 3.86%, 4.14%를 나타냈다. STRO-1은 35.62%로 발현되었다. 이상의 결과에서 과잉치 치수 유래 세포는 간엽성 줄기세포의 특성을 가지며, 3계대와 10계대 모두에서 간엽성 줄기세포의 특성을 유지하고 있다고 사료된다. 이에 빠른 성장 속도와 늦은 계대까지 유지되는 줄기세포능을 고려할 때, 치아 유래 줄기세포의 공여부로서 매복 과잉치의 충분한 활용 가능성을 확인하였다.

성견 치주인대세포의 줄기세포 특성 연구 (Stem cell properties of cells derived from canine periodontal ligament)

  • 김경화;김수환;설양조;이용무
    • Journal of Periodontal and Implant Science
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    • 제37권3호
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    • pp.479-488
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    • 2007
  • In spite of the attention given to the study of mesenchymal stem cells derived periodontal ligament (PDL), there is a lack of information about canine PDL cells. In this study, we characterized canine PDL cells to clarify their stem cell properties, including self renewal, proliferate rate, stem cell markers and multipotency. PDL cells were obtained from extracted premolars of canines, following a colony forming assay and proliferation rate of sub-confluent cultures of cells for self-renewal, immunostaining for STRO-1 and CD146/MUC18 and a differentiation assay for multipotency. Canine PDL cells formed single-cells colonies and 25% of the PDL cells displayed positive staining for BrdU. The cells expressed the mesenchymal stem-cell markers, STRO-1 and CD146/MUC18. Under defined culture conditions, the cells differentiated into osteoblasts and adipocytes, but the cells didn't differentiated into chondrocytes. The findings of this study indicated that the canine PDL cells possess crucial stem cells properties, such as self-renewal and multipotency, and express the mesenchymal stem cell markers on their surface. The isolation and characterization of canine PDL cells makes it feasible to pursue preclinical models of periodontal regeneration in canine.

Gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow

  • Kim, Su-Hwan;Kim, Young-Sung;Lee, Su-Yeon;Kim, Kyoung-Hwa;Lee, Yong-Moo;Kim, Won-Kyung;Lee, Young-Kyoo
    • Journal of Periodontal and Implant Science
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    • 제41권4호
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    • pp.192-200
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    • 2011
  • Purpose: The aim of this study is to compare the gene expression profile in mesenchymal stem cells derived from dental tissues and bone marrow for characterization of dental stem cells. Methods: We employed GeneChip analysis to the expression levels of approximately 32,321 kinds of transcripts in 5 samples of bone-marrow-derived mesenchymal stem cells (BMSCs) (n=1), periodontal ligament stem cells (PDLSCs) (n=2), and dental pulp stem cells (DPSCs) (n=2). Each cell was sorted by a FACS Vantage Sorter using immunocytochemical staining of the early mesenchymal stem cell surface marker STRO-1 before the microarray analysis. Results: We identified 379 up-regulated and 133 down-regulated transcripts in BMSCs, 68 up-regulated and 64 down-regulated transcripts in PDLSCs, and 218 up-regulated and 231 down-regulated transcripts in DPSCs. In addition, anatomical structure development and anatomical structure morphogenesis gene ontology (GO) terms were over-represented in all three different mesenchymal stem cells and GO terms related to blood vessels, and neurons were over-represented only in DPSCs. Conclusions: This study demonstrated the genome-wide gene expression patterns of STRO-$1^+$ mesenchymal stem cells derived from dental tissues and bone marrow. The differences among the expression profiles of BMSCs, PDLSCs, and DPSCs were shown, and 999 candidate genes were found to be definitely up- or down-regulated. In addition, GOstat analyses of regulated gene products provided over-represented GO classes. These data provide a first step for discovering molecules key to the characteristics of dental stem cells.

Differentiation and characteristics of undifferentiated mesenchymal stem cells originating from adult premolar periodontal ligaments

  • Kim, Seong Sik;Kwon, Dae-Woo;Im, Insook;Kim, Yong-Deok;Hwang, Dae-Seok;Holliday, L. Shannon;Donatelli, Richard E.;Son, Woo-Sung;Jun, Eun-Sook
    • 대한치과교정학회지
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    • 제42권6호
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    • pp.307-317
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    • 2012
  • Objective: The purpose of this study was to investigate the isolation and characterization of multipotent human periodontal ligament (PDL) stem cells and to assess their ability to differentiate into bone, cartilage, and adipose tissue. Methods: PDL stem cells were isolated from 7 extracted human premolar teeth. Human PDL cells were expanded in culture, stained using anti-CD29, -CD34, -CD44, and -STRO-1 antibodies, and sorted by fluorescent activated cell sorting (FACS). Gingival fibroblasts (GFs) served as a positive control. PDL stem cells and GFs were cultured using standard conditions conducive for osteogenic, chondrogenic, or adipogenic differentiation. Results: An average of $152.8{\pm}27.6$ colony-forming units was present at day 7 in cultures of PDL stem cells. At day 4, PDL stem cells exhibited a significant increase in proliferation (p < 0.05), reaching nearly double the proliferation rate of GFs. About $5.6{\pm}4.5%$ of cells in human PDL tissues were strongly STRO-1-positive. In osteogenic cultures, calcium nodules were observed by day 21 in PDL stem cells, which showed more intense calcium staining than GF cultures. In adipogenic cultures, both cell populations showed positive Oil Red O staining by day 21. Additionally, in chondrogenic cultures, PDL stem cells expressed collagen type II by day 21. Conclusions: The PDL contains multipotent stem cells that have the potential to differentiate into osteoblasts, chondrocytes, and adipocytes. This adult PDL stem cell population can be utilized as potential sources of PDL in tissue engineering applications.

Osteogenic Potency of Nacre on Human Mesenchymal Stem Cells

  • Green, David W.;Kwon, Hyuk-Jae;Jung, Han-Sung
    • Molecules and Cells
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    • 제38권3호
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    • pp.267-272
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    • 2015
  • Nacre seashell is a natural osteoinductive biomaterial with strong effects on osteoprogenitors, osteoblasts, and osteoclasts during bone tissue formation and morphogenesis. Although nacre has shown, in one study, to induce bridging of new bone across large non-union bone defects in 8 individual human patients, there have been no succeeding human surgical studies to confirm this outstanding potency. But the molecular mechanisms associated with nacre osteoinduction and the influence on bone marrow-derived mesenchymal stem cells (BMSC's), skeletal stem cells or bone marrow stromal cells remain elusive. In this study we highlight the phenotypic and biochemical effects of Pinctada maxima nacre chips and the global nacre soluble protein matrix (SPM) on primary human bone marrow-derived stromal cells (hBMSCs) in vitro. In static co-culture with nacre chips, the hBMSCs secreted Alkaline phosphatase (ALP) at levels that exceeded bone morphogenetic protein (rhBMP-2) treatment. Concentrated preparation of SPM applied to Stro-1 selected hBMSC's led to rapid ALP secretions, at concentrations exceeding the untreated controls even in osteogenic conditions. Within 21 days the same population of Stro-1 selected hBMSCs proliferated and secreted collagens I-IV, indicating the premature onset of an osteoblast phenotype. The same SPM was found to promote unselected hBMSC differentiation with osteocalcin detected at 7 days, and proliferation increased at 7 days in a dose-dependent manner. In conclusion, nacre particles and nacre SPM induced the early stages of human bone cell differentiation, indicating that they may be promising soluble factors with osteoinductive capacity in primary human bone cell progenitors such as, hBMSC's.

협부지방에서 성체 줄기세포의 분리와 골모 세포로의 분화 (DIFFERENTIATION OF ADULT STEM CELL DERIVED FROM BUCCAL FAT PAD INTO OSTEOBLAST)

  • 표성운;박장우;이일규;김창현
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제32권6호
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    • pp.524-529
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    • 2006
  • For the repairing of bone defect, autogenous or allogenic bone grafting remains the standard. However, these methods have numerous disadvantages including limited amount, donor site morbidity and spread of diseases. Tissue engineering technique by culturing stem cells may allow for a smart solution for this problem. Adipose tissue contains mesenchymal stem cells that can be differentiate into bone, cartilage, fat or muscle by exposing them to specific growth conditions. In this study, the authors procured the stem cell from buccal fat pad and differentiate them into osteoblast and are to examine the bone induction capacity. Buccal fat-derived cells (BFDC) were obtained from human buccal fat pad and cultured. BFDC were analyzed for presence of stem cell by immunofluorescent staining against CD-34, CD-105 and STRO-1. After BFDC were differentiated in osteogenic medium for three passages, their ability to differentiate into osteogenic pathway were checked by alkaline phosphatase (ALP) staining, Alizarin red staining and RT-PCR for osteocalcin (OC) gene expression. Immunofluorescent and biochemical assays demonstrated that BFDC might be a distinguished stem cells and mineralization was accompanied by increased activity or expression of ALP and OC. And calcium phosphate deposition was also detected in their extracelluar matrix. The current study supports the presence of stem cells within the buccal fat pad and the potential implications for human bone tissue engineering for maxillofacial reconstruction.

BMP-2에 의한 협부 지방 성체 줄기세포의 골형성 (OSTEOGENESIS BY BMP-2 IN ADULT STEM CELL DERIVED FROM BUCCAL FAT PAD)

  • 김창현;박철헌;이일규;표성운
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제34권4호
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    • pp.412-418
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    • 2008
  • Bone morphogenetic proteins (BMPs) in combination with stem cells gain more significance for their use in bone tissue engineering. The mesenchymal stem cell can be differentiated into osteoblast by the treatment of BMP. The aim of this study is to characterize the osteogenic differentiation process of adult stem cells derived from buccal fat pad according to BMP-2 within culture media and decide the appropriate concentration of BMP-2 to facilitate osteogenesis. The authors procured the stem cell from buccal fat pad and analyzed for presence of stem cell by flow cytomety against CD-34, CD-105 and STRO-1. The buccal fat derived stem cells (BFDC) were treated by application of the different concentration with BMP-2 of 0, 10, 50, 100 and 200ng/ml, respectively. And their ability to differentiate into osteogenic pathway were checked by alkaline phosphatase(ALP) staining, Alizarin red staining and RT-PCR for osteocalcin(OC) gene expression at 7, 14 and 21day of culture. Flow cytometric analysis and biochemical assays demonstrated that BFDC might be a distinguished stem cells, and mineralization was accompanied in proportion to BMP-2 concentration. However, with 100ng/ml concentration of BMP-2, the BFDC demonstrated most efficient staining pattern of ALP and Alizarin red. The feasibility of the osteogenic differentiation in the group of both 50ng/ml and 200ng/ml of BMP-2 showed similar activity and relatively weaker than that of 100ng/ml. These results suggest that the BMP-2 stimulate osteogenesis by BFDC effectively and that bone induction might be controlled through negative regulatory feedback in higher concentration.

성견 치계줄기세포 및 골수줄기세포 특성에 관한 연구 (Investigation of postnatal stem cells from canine dental tissue and bone marrow)

  • 진민주;김영성;김수환;김경화;이철우;구기태;김태일;설양조;구영;류인철;정종평;이용무
    • Journal of Periodontal and Implant Science
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    • 제39권2호
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    • pp.119-128
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    • 2009
  • Purpose: The aim of this study was to evaluate the stemness of cells from canine dental tissues and bone marrow. Methods: Canine periodontal ligament stem cells (PDLSC), alveolar bone stem cells (ABSC) and bone marrow stem cells(BMSC) were isolated and cultured. Cell differentiations (osteogenic, adipogenic and chondrogenic) and surface antigens (CD146, STRO-1, CD44, CD90, CD45, CD34) were evaluated in vitro. The cells were transplanted into the subcutaneous space of nude mice to assess capacity for ectopic bone formation at 8 weeks after implantation. Results: PDLSC, ABSC and BMSC differentiated into osteoblasts, adipocytes and chondrocytes under defined condition. The cells expressed the mesenchymal stem cell markers differently. When transplanted into athymic nude mice, these three kinds of cells with hydroxyapatite /${\beta}$- tricalcium phosphate (HA/TCP) carrier showed ectopic bone formation. Conclusions: This study demonstrated that canine dental stem cells have stemness like bone marrow stem cells. Transplantation of these cells might be used as a therapeutic approach for dental stem cell-mediated periodontal tissue regeneration.

A Study on the Meaning and Strategy of Keyword Advertising Marketing

  • Park, Nam Goo
    • 유통과학연구
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    • 제8권3호
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    • pp.49-56
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    • 2010
  • At the initial stage of Internet advertising, banner advertising came into fashion. As the Internet developed into a central part of daily lives and the competition in the on-line advertising market was getting fierce, there was not enough space for banner advertising, which rushed to portal sites only. All these factors was responsible for an upsurge in advertising prices. Consequently, the high-cost and low-efficiency problems with banner advertising were raised, which led to an emergence of keyword advertising as a new type of Internet advertising to replace its predecessor. In the beginning of 2000s, when Internet advertising came to be activated, display advertisement including banner advertising dominated the Net. However, display advertising showed signs of gradual decline, and registered minus growth in the year 2009, whereas keyword advertising showed rapid growth and started to outdo display advertising as of the year 2005. Keyword advertising refers to the advertising technique that exposes relevant advertisements on the top of research sites when one searches for a keyword. Instead of exposing advertisements to unspecified individuals like banner advertising, keyword advertising, or targeted advertising technique, shows advertisements only when customers search for a desired keyword so that only highly prospective customers are given a chance to see them. In this context, it is also referred to as search advertising. It is regarded as more aggressive advertising with a high hit rate than previous advertising in that, instead of the seller discovering customers and running an advertisement for them like TV, radios or banner advertising, it exposes advertisements to visiting customers. Keyword advertising makes it possible for a company to seek publicity on line simply by making use of a single word and to achieve a maximum of efficiency at a minimum cost. The strong point of keyword advertising is that customers are allowed to directly contact the products in question through its more efficient advertising when compared to the advertisements of mass media such as TV and radio, etc. The weak point of keyword advertising is that a company should have its advertisement registered on each and every portal site and finds it hard to exercise substantial supervision over its advertisement, there being a possibility of its advertising expenses exceeding its profits. Keyword advertising severs as the most appropriate methods of advertising for the sales and publicity of small and medium enterprises which are in need of a maximum of advertising effect at a low advertising cost. At present, keyword advertising is divided into CPC advertising and CPM advertising. The former is known as the most efficient technique, which is also referred to as advertising based on the meter rate system; A company is supposed to pay for the number of clicks on a searched keyword which users have searched. This is representatively adopted by Overture, Google's Adwords, Naver's Clickchoice, and Daum's Clicks, etc. CPM advertising is dependent upon the flat rate payment system, making a company pay for its advertisement on the basis of the number of exposure, not on the basis of the number of clicks. This method fixes a price for advertisement on the basis of 1,000-time exposure, and is mainly adopted by Naver's Timechoice, Daum's Speciallink, and Nate's Speedup, etc, At present, the CPC method is most frequently adopted. The weak point of the CPC method is that advertising cost can rise through constant clicks from the same IP. If a company makes good use of strategies for maximizing the strong points of keyword advertising and complementing its weak points, it is highly likely to turn its visitors into prospective customers. Accordingly, an advertiser should make an analysis of customers' behavior and approach them in a variety of ways, trying hard to find out what they want. With this in mind, her or she has to put multiple keywords into use when running for ads. When he or she first runs an ad, he or she should first give priority to which keyword to select. The advertiser should consider how many individuals using a search engine will click the keyword in question and how much money he or she has to pay for the advertisement. As the popular keywords that the users of search engines are frequently using are expensive in terms of a unit cost per click, the advertisers without much money for advertising at the initial phrase should pay attention to detailed keywords suitable to their budget. Detailed keywords are also referred to as peripheral keywords or extension keywords, which can be called a combination of major keywords. Most keywords are in the form of texts. The biggest strong point of text-based advertising is that it looks like search results, causing little antipathy to it. But it fails to attract much attention because of the fact that most keyword advertising is in the form of texts. Image-embedded advertising is easy to notice due to images, but it is exposed on the lower part of a web page and regarded as an advertisement, which leads to a low click through rate. However, its strong point is that its prices are lower than those of text-based advertising. If a company owns a logo or a product that is easy enough for people to recognize, the company is well advised to make good use of image-embedded advertising so as to attract Internet users' attention. Advertisers should make an analysis of their logos and examine customers' responses based on the events of sites in question and the composition of products as a vehicle for monitoring their behavior in detail. Besides, keyword advertising allows them to analyze the advertising effects of exposed keywords through the analysis of logos. The logo analysis refers to a close analysis of the current situation of a site by making an analysis of information about visitors on the basis of the analysis of the number of visitors and page view, and that of cookie values. It is in the log files generated through each Web server that a user's IP, used pages, the time when he or she uses it, and cookie values are stored. The log files contain a huge amount of data. As it is almost impossible to make a direct analysis of these log files, one is supposed to make an analysis of them by using solutions for a log analysis. The generic information that can be extracted from tools for each logo analysis includes the number of viewing the total pages, the number of average page view per day, the number of basic page view, the number of page view per visit, the total number of hits, the number of average hits per day, the number of hits per visit, the number of visits, the number of average visits per day, the net number of visitors, average visitors per day, one-time visitors, visitors who have come more than twice, and average using hours, etc. These sites are deemed to be useful for utilizing data for the analysis of the situation and current status of rival companies as well as benchmarking. As keyword advertising exposes advertisements exclusively on search-result pages, competition among advertisers attempting to preoccupy popular keywords is very fierce. Some portal sites keep on giving priority to the existing advertisers, whereas others provide chances to purchase keywords in question to all the advertisers after the advertising contract is over. If an advertiser tries to rely on keywords sensitive to seasons and timeliness in case of sites providing priority to the established advertisers, he or she may as well make a purchase of a vacant place for advertising lest he or she should miss appropriate timing for advertising. However, Naver doesn't provide priority to the existing advertisers as far as all the keyword advertisements are concerned. In this case, one can preoccupy keywords if he or she enters into a contract after confirming the contract period for advertising. This study is designed to take a look at marketing for keyword advertising and to present effective strategies for keyword advertising marketing. At present, the Korean CPC advertising market is virtually monopolized by Overture. Its strong points are that Overture is based on the CPC charging model and that advertisements are registered on the top of the most representative portal sites in Korea. These advantages serve as the most appropriate medium for small and medium enterprises to use. However, the CPC method of Overture has its weak points, too. That is, the CPC method is not the only perfect advertising model among the search advertisements in the on-line market. So it is absolutely necessary that small and medium enterprises including independent shopping malls should complement the weaknesses of the CPC method and make good use of strategies for maximizing its strengths so as to increase their sales and to create a point of contact with customers.

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