커피나무(Coffea arabica L. ‘Blue Mountain’)의 분화 관상식물로 개발할 목적으로 적정 용토와 액비농 도를 구명하기 위한 실험을 수행하였다. 분용토 선발을 위하여 토실이상토, 폐암면입자 + 일반상토, 폐암면입 자 + PUR(스펀지)입자, 폐암면입자 + 목재칩, 그리고 일반상토를 이용하여 약 3개월간 재배한 결과, 초장 및 수관폭 등 생육지표에서 폐암면입자 + 일반상토와 폐암면입자 + PUR입자가 가장 좋았다. 엽록소와 엽색 에서 토실이상토, 폐암면입자 + 일반상토, 그리고 폐암 면입자 + PUR입자에서는 진한 녹색으로 관상가치가 높았으나, 폐암면입자 + 목재칩과 일반상토구에서는 녹 색이 엷어져 상품성이 많이 떨어졌다. 적정 액비 농도 를 구명하기 위해 Hyponex를 2000, 1000, 500배 희석액으로 1회/주 살포한 결과, 1000배 희석액까지는 뚜렷한 차이가 보이지 않았으나 500배 희석액에서는 생육이 더 좋고 엽색도 진한 녹색으로 상품가치가 더 좋은 것으로 나타났다. 결론적으로 폐암면입자에 일반 상토나 PUR입자를 혼합하여 사용하고 Hyponex 500 배 희석액을 주 1회 관주하면 좋은 품질의 커피나무 분화를 생산할 수 있을 것으로 판단되었다.

A case of a distinctive variety of basalαd squamous cell carcinoma (B8CC) of the floor of mouth is described Histologic evaluation of the tumor showed lobules and aggregates of medium-sized basaloid cells with distinctive periph eral palisading and focal areas of comedo- necrosis. This appearance together with microcystic spaces simulated that of an adenoid cystic carcinoma. Accompanying epithelial dysplasia of the overlying mucosa was found. Immunohistochemistry of the tumor revealed diffuse strong expression for cytokeratin AE1/3, focally positive for Epithelial Membrane Antigen in the inner cells of tubular structures However, CEA, 8-100, smooth muscle actin, and vimentin were negative. The histologic differential diagnosis considered were adenoid cystic carcinoma and adenosquamous cell carcinoma Immunohistochemistry and awareness of this unusual pattern of B8CC will facilitate the correct diagnosis being reached.

The purpose of this study was to examine the effects of vi tamin D3 and 1'etinoic acid(RA) on the human mesenchymal stem ce!ls(MSC) g1'owth and osteogenic differentiations. Cell proliferation, mineralization, cell cycle, expression of cell cycle regu l atOJγ proteins and markers fo1' osteogenic differenatiaiton were determined by MTI assay, mineralization assay, flow cytomet1'Y‘ and Western blot analysis, respectively. Cell viability was dec1'ease by each vitamin D3 and RA added to MSC. it was more decrease by vitamin D3 and RA. Mineralized nodule formation revealed similar expression pattern with positive cont rol group at vitamin D3 and RA mixed add to MSC. At vitamin D3 and RA mixed add to MSC after 7 days of incubation was increase G1 s tage. after 21 days of incubation was inhibit cell cycle prog1'ess by inc1'ease of sub-G1 Treatment vitamin D3 to MSC inhibits p53 and p21, but inc1'ease pRb. RA inhibit p53, but increase p21 and pRb, vitamin D3 plus RA group was same as added RA group. so two vitamin was effect to inhibited cell growth each different mechanism. Expression of BMP-2 protein was prominent in osteogonic supplement treated g1'oup of MSC at 2 weeks cultivation days, but vi tamin D3 treatment decreased BMP-2 expression rather than in (+) control group. BSP protein was notably increased in the OS compa red to positive controls at 2 weeks cultivation, but similar to that of vitamin D3 group t1'eatment group and was least expressed in plus RA mixed group, at 3 weeks, BSP expression was similar to 1'esult of 2 weeks Collectively, these results shows that vitamin D3 and RA have diffe1'ential effects on the MSCs g1'owth and differ entia tion 211

Dentin is a mineralized tissue formed by odontoblasts that are differentiated from ectomesenchymal cells , The molecul ar mech anism of odontoblast diffe rentiation remains unclear, Amino acid transporters play an important role in s up plying nutri tion to normal a nd ca ncer cells including odntoblasts, and for cell proliferation , Amino acid transport system L is a maj or nutrient t ransport system responsible for the Na+' -independent transport o[ neutral amino acids incJuding several essentiaJ amino acids , The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2) , In this study, the expression pattern and role of amino acid transport system L were, therefore, investigated in the differentiation of MDPC-23 cells derived from mouse dental papilla celJs , To determi ne the expression Jevel o[ amino acid transport system L participating in intracelJ ular transport of amino acids in the differentiat ion 0 1' MDPC-23 cells, it was examined by RT-PCR, observation of cell morphoJogy‘ A1izaline red-S staining ancl uptake analysis after inclucing experimental differentiation in MDPC-23 cells The res ults were as follows , The LAT1 mRNA was expressed in the early stage of MDPC-23 cell differentiation , The expression leveJ was gradually increased by time course and it was decreased after the late stage, The LAT2 mRNA was not observed in the earJy stage of MDPC-23 cell differentiation, The LAT2 mRNA was expressed at the 11 days 0 1' MDPC-23 cell differentiation and the expression level was gradually decreased by time course, There was no changes in the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during the differentiation of MDPC-23 cells , The expression of ON mRNA was graduaJJy decreased but the expression of ALP mRNA was increased during differentiation of MDPC-23 cells , The L-Ieucine uptake was increased by time cour se from the early stage to the 9 days in MDPC-23 cell differentiation , The amount of L-Ieucine uptake was maintained to the 11 and 14 days of MDPC-23 cell differentiation As the resul ts‘ it is considered that among neutral amino acid transport system L in differentiation of MDPC-23 cells , the LATl has a key role in cell proliferation in the early stage and middle stage of cell differentiation and the LAT2 has an important roJe in ceJJ differenti ation and mineralization in the Jate stage of cell differentiation for providing cells with neutral a mino acids incJuding several essentiaJ amino acids

NOD/SCID 마우스는 선천성 면역결핍을 지닌 마우스로서 이종 세포 및 조직 이식을 위한 실험동물로서 가장 많이 활용되고 있다. 본 연구는 돼지의 골수조직에서 채취한 조혈줄기세포를 면역결핍마우스의 정맥 주입을 통하여 생체 내 주입을 실시한 결과, 마우스의 조혈조직에서 대단히 높은 돼지 T 면역세포의 증식이 관찰되었다. 유세포 분석기를 이용해 돼지 골수 조혈세포 생체 이식 6주의 마우스에서의 돼지 T 면역세포의 증식과 분화 특성을 분석한 결과, 마우스 조혈조직인 골수(5.4±1.9%), 비장(15.4±7.3%), 간(21.3±1.4%), 림프절(33.5±32.8%)에서 돼지 조혈줄기세포 유래 T 세포의 증식과 분화가 관찰되었고, 돼지 helper T 세포와 cytotoxic T 세포의 발달도 확인되었다. 또한 조직 면역염색을 통하여 마우스의 비장조직에 이식한 돼지 면역세포의 증식을 관찰하였다. 본 연구는 NOD/SCID 마우스를 이용해 돼지 조혈줄기세포로부터 T 면역세포로의 분화 및 발달과정을 생체 내에서 분석할 수 있는 유용한 동물모델로서 이용할 수 있음을 보여준다.

Previous ly we have s hown that fï brob last• growth factor-2 (FGF-2) and dexamethasone (Dex) in combination strongly stimulate both p l 이 i fe rati o n a nd differe nt iation of mesenchymal stem cells (MSCs) into osteoblasts and adipocytes, In the present s tudy we invesL igaLed whether inhibition 01' FGF-2 and Dex-induced adipogenic differentiation of bone marrow derived s Lem cells (BMSCs) by GW9662, an antagoni s t of proxisome proliferators-activated receptol γ (PPARy) which plays a key role in ad ipogenic differentiation , enhances proliferation and osteoblastic differentiation of BMSCs Proliferation 01' BMSCs t reated wi 네 FGF-2 a nd Dex was further increased by GW9662 up to 9,7, 10,6, and 7,2% at 3, 5, and 7 days of cul Lu re , Expansion of BMSCs with FGF-2, Dex and GW9662 followed by osteoblastic different iation showed that osteoblas tic differentiation 01' BMSCs was in creased by 37 % (p=O, 01) compared to those expanded with FGF-2 and Dex, ln contrast , ad i pogenic di fferenti a tion of FGF-2 and Dex-expanded BMSCs was substantially reduced to 14% (p=O, 036) by GW9662, Taken toget her , these resul ts demonstrate that FGF-2 and Dex in combination with GW9662 f ur t her stimu late proliferation 01' BMSCs and those cells expanded with these factors acquire enhanced potentiaIs to be dif ferentiated i n to osteoblas ts

Extensive oral mucosa loss from a variety of conditions is associated with significant functional morbidity and mortality. Although it is known that keratinocytes are a rich source of wound healing promoting factors such as transforming growth factor-β1(TGF-β1), it is not clear whether differentiated keratinocytes in a multi-layer form release this multi-functional growth factor. This study examined the hypothesis that keratinocytes in mono- and multi-layer forms expressed different levels of TGF-β1. When NHOK reached confluency in serum free medium(KBM), in test medium containing 1.2 mM Ca++ KBM NHOK were allowed to form multi-layers and differentiate. The purpose of this study were to investigate the mRNA level of TGF-β1, FGF-2, and TIMP-1 by RT-PCR analysis and also to evaluate the expression of TGF-β1 and involucrin in keratinocytes at different times of the onset of differentiation. The numbers and sizes of these nodules were increased as the process of keratinocyte differentiation proceed. Cultured NHOK in preconfluency under KBM medium expressed a significantly higher level of TGF-β1 relative to those grown in multi-layer forms, while the level of TGF-β1 mRNA gradually reduced to its lowest level at 7 days of growing cells in test medium. Cultured NHOK in preconfluency of KBM medium expressed a lower level of FGF-2 and TIMP-1 relative to those grown in multi-layer forms, while the level of FGF-2 and TIMP-1 mRNA showed the highest level at 3 days at gradually reduced to its lowest level at 7 days of growing cells in test medium. As a differentiation marker for keratinocytes at different time points, the highest level of involucrin mRNA expression was found at the later stage of cell differentiation. It suggested that the expression of TGF-β1 mRNA be consistent with the expression of FGF-2 and TIMP-1 mRNA in NHOK grown in high calcium medium during the terminal differentiation. But differentiated NHOK expressing higher involucrin mRNA could show constant espression of TGF-β1, FGF-2 and TIMP-1.

Chicory(Cichorium intybus L.)의 우량 종근 생산을 위해 파종 시기를 1개월 간격으로 하여 종근 형성 및 화아분화에 미치는 영향을 조사하였다. 화아분화 개시는 4월 파종은 6월 17일, 5월 파종은 7월 2일, 6월 파종은 7월 30일, 7월 파종은 10월 1일에 각각 화아가 분화되었으나 8월과 9월 파종은 전혀 분화되지 않았다. 파종시기별 생육특성에서 4~7월 파종은 종근의 무게가 150g에 도달하기 전에 화아가 분화되어 종근을 이용할 수가 없었으며, 9월 파종은 생육동안 외기온도가 낮아 겨울을 넘기면서 종근이 정상적으로 발육하지 못하는 결과를 보였고, 8월 파종은 파종 후 약 90일이 되는 시기에 Chicon을 생산할 수 있는 종근의 적절한 중량인 200g이상의 이상적인 굴취기에 도달하였으며, 생육기간동안 화아분화가 일어나지 않았다. 따라서 우리나라 기후에 따른 우량 Chicory 종근을 생산하기 위한 적정 파종기는 8월인 것으로 조사되었다.

폐펄라이트와 폐유리를 가공하여 입상화한 cellular glass foam(CGF)을 각각 입상암면과 피트모스를 혼합한 배지에서 매트저면관수한 Viola×wittrockiana 'Majestic GT Scarlet Shadow'를 재배하여 개화와 생육을 조사하였다. 엽수, 분지수, 지상부 생체중과 건물중, 엽면적과 총엽록소 함량은 폐펄라이트+입상암면+피트모스 (25:50:25, v/v/v) 혼합배지에서 가장 좋았으며, 초장과 최대근장은 CGF+피트모스(25:75, v/v) 혼합배지에서 가장 좋았다. 대조구인 폐펄라이트+피트모스(25:75, v/v) 혼합배지와 비교하여 CGF+피트모스(25:75, v/v)에서의 초장, 엽수, 근장, 총엽록소 함량, 지상부와 지하부의 생체중과 건물중이 더 켰다. 그러나 폐펄라이트+입상암면(25:75, v/v) 혼합배지와 비교하여 CGF+입상암면(25:75, v/v)에서의 생육은 매우 저조하였다.

본 연구는 단위발생유래 생쥐 배아줄기세포(P-mES)지가 체외수정유래 생쥐 배아줄기세포 (mES)와 마찬가지로 기능성 심근세포로 체외 분화되는지를 조사하였다. 각 세포주 P-mES04와 MES03를 4일간 부유 배양하여 배아체 (EB)를 형성한 다음 4일간 DMSO를 추가적으로 처리한 뒤 젤라틴이 코팅된 배양접시에 부착시켰다(4-/4+). P-mES04와 mES03으로부터 수축성 심근세포 생성 여부를 30일간 관찰한 결과, 각각 13일(69.83%)과 22일 (61.3%)에 누적 형성율이 가장 높았다. 면역 세포화학염색 결과, 수축성을 나타내는 P-mES04 세포는 수축성 mES03 세포에서와 같이 근육 특이적인 anti-sarcomeric a-actinin 항체와 심근 특이적인 anti-cardiac troponin I 항체에 염색되는 것을 확인하였다. 또한 RT-PCR 결과, 수축성을 나타내는 P-mES04 세포는 심근특이적인 L-type calcium channel, a1C, cardiac myosin heavy chain a, cardiac muscle heavy polypeptide 7β, GATA binding protein 4와 atrial natriuretic factor는 발현하나, 골격근 특이적인 L-type calcium channel, a1S는 발현하지 않아 웅성 성체의 심장세포와 유사한 양상을 보였다. 본 연구의 결과는 단위발생 유래 생쥐 배아 줄기세포를 배아줄기세포의 연구의 대체제로 이용할 수 있음을 보여준다.

It is well kwon that HPV have been strongly linked to progression of or al squamous cell carcinoma‘ Effici ent im mortalization of nonnal human oral keratinocyte(NHOK) should provid further evidence for the role HPV in tumorogenes is ‘ Because IHOK(I mmortali zed human oral keratinocyte) has been considered as a moclel syst em for study ing I-!PV- linkecl oral ca rcinogenes is , it is important to pursue the differenti ati al change of IHOK cul t ure moclel during t he culture passage, The purposes of this study were to examine the cha ra r’ ct eristic clifferential changes of cul turecl immorta lizecl human ora l keratinocytes during long term passage, and to apply these results to or al carcinogenes is in the future, NI-!OK was primarily incubated at 370C and 5% C02 under KBM bullet kJt IHOK was co ntinuously cul t ured towarcl 100th passage(two times per week) , Growth curve of NI-!OK and II-!OK clepend on clùture passage was taken For examining the cha racte ri s t ic clifferential changes of II-!OK, transrnission electron microscope, 1ì'ansgluta miase activity‘ E6/E7 mRNA detect ion, a ncl tumorogenecity were done 10th II-!OK showecl sl ight polygonal flattencl cells and sometimes apoptotic cells ‘ while 100th IHOK showecl increased polygonal cell s ‘ Cultu recl 100th IHOK showed r ela tively resis tant growth to high calcium than 10th II-!OK Microvilli from 10th II-!OK was not connect ecl with each other, ancl scatte red cytokeratin fil aments of 10th II-IOK. while decreased cytokeratin filaments in cytoplasm & prominent clesmosome of 100th IHOK. During the terminal differ entiation in cultured IHOK, induction of TGase 1 activity of 10th II-!OK was higher than that of100th IHOK mRNA E6E7 expresson was cletected and unchangable in both cul tured cells There was no tumorogenecity inclucecl by both culturecl cel ls. Although late passage IHOK showecl less r esemblance to NHOK, and lower TGase 1 acti vi ty than ea rly passage IHOK, it suggested that these cells should be 110t yet fully differ entiatecl to oral squ a mous ce ll carcinoma cells

고품질 분화생산을 위한 저면관수 시스템인 C-형강 매트재배 시 수용성 비료와 완효성 비료의 농도에 따른 카네이션의 생육을 연구하기 위하여 본 실험을 수행하였다. 수용성 비료를 0.8~1.0 g·L-1 처리한 T08 처리구에서 생체중, 건물중, 엽수, 엽면적, 초장, 분지수의 생육량에서 가장 양호하였다. 생체중, 건물중, 및 초장의 증가에 있어서 증가량에서의 차이는 있었지만 생육 60일을 전후로 비료의 종류에 관계없이 증가량이 늘어났다. 특히 총 엽면적은 경우 생육 90일을 전후로 증가가 완만해지는 경향을 나타내었다. 이 결과를 통하여 C-형강 매트 재배시스템에서 분화용 카네이션을 재배할 경우 수용성 비료를 공급하는 것이 완효성 비료를 공급하는 것보다 효과적인 것으로 나타났고, 수용성 비료의 농도는 0.8~1.0 g·L-1이 카네이션의 재배에 적정한 농도로 판명 되었다.

Nuclear factor 1 (NFI) was discovered as a protein required for adenovirus DNA replication in vitro, but it is now clear that NFI protein plays an important role in the expression of many cellular genes. NFI-C null mice demonstrated aberrant odontoblast differentiation, abnormal dentin formation, and thus molar lacking roots while other tissues/or gans in the body, including ameloblasts appear to be unaffected and normal. However, little is known about the mechanism of NFI -C function in odontoblast differentiation and dentin formation. In this study, in order to elucidate the molecular mechanisms of odntoblast differentiation, we examined morphological characteristics of the aberrant odontoblast in NFI-C null mice. we also evaluate the expression of dentin sialophosphoprotein (DSPP) and bone sialoprotein (BSP) mRNAs in the MDPC-23 cells by northern analysis after over-expression and inactiγation of NFI -C into mouse MDPC-23 cells Odontoblasts of the NFI-C null mouse were round in shape, lost their polarity, organized as a sheet of cells, and trapped in osteodentin-like mineralized tissue. Abnormal odontoblasts of NFI-C null mouse revealed the absence of an intercellular junctional complex known as the t erminal webs. MDPC-23 cells started to express DSPP mRNA beginning from the postnatal day of 14 and showed a steady increase as differentiating into odontoblasts. Over-expression of NFI -C increased the expression of DSPP mRNA. Inactivation of NFI - C induced BSP mRNA expression. These results suggest that NFI-C plays an important role in odontoblast differentiation in a cell-type specific manner and thus in dentin formation

Periodontalligament (PDL) fibroblasts have an ectomesenchymal origin and are known to participate not only in formation of PDL but also in the repair and regeneration of the a이acent alveolar bone and cementum. However, little is known about the molecular mechanism which is related to the development and differentiation of PDL cells. Recendy, we reported the PDLs (a periodontalligament-specific) 22 as a PDL fibroblast-specific mRNA which is not expressed in gingival fibroblasts. In this study, to examine the expression and functional characterization of PDμ22 mRNA and prαein in development and differentiation of periodontal 따sue ， we carried out northem analysis, insitu hybridization, immunofluorescence and immunohistochemistry. The expression of PDLs22 mRNA was increased with PDL cell differentiation from the confluent to multilayer stage but decreased slighdy with mineralized nodule formation in vitro. πle PDLs22 protein was localized on the nuclear membrane and expressed throughout the differentiation of PDL fibroblasts in vitro. The PDLs22 mRNA and protein were expressed in the differentiating cementoblasts, PDL fibroblasts and osteoblasts along the r∞t surface and alveolar bone of the developing rat teeth. These results indicate that the PDLs22 plays an irnportant role in the differentiation of cementoblasts and osteoblasts and thus homeostasis of cementum, PDL and alveolar bone.