Embryoid bodies (EBs) generated from human embryonic stem cells (hESCs) include spontaneously induced endodermal lineage cells (ELCs). Activin-A plays important roles in the endoderm differentiation of hESCs. Despite studies on the generation of ELCs from hESCs with treatment of Actvin-A, it was unclear for localization and pattern of ELCs by Activin-A during differentiation of hESCs. Accordingly in this study, we knew that Actvin-A increased the cystic EBs formation, including the highly enriched AFP (endoderm lineage specific marker)-expressing cells in the surface of cystic EBs. To induce the EBs formation from undifferentiated hESCs, cells were transferred onto petri-dish and cultured in suspension condition without bFGF removed hESC media (EB media) for 3 days. Next to investigate the effect of Activin-A, EBs were subsequently cultured in EB media supplement with 100 ng/ml Activin-A for 3 days. After 5～7 days of Activin-A treatment, cystic EBs began to appear which increased in numbers reaching ～60% of initially formed EBs over 5 days. Endoderm lineage marker, AFP were highly expressed and specifically localized at the surface region of cystic EBs comparison with normal EBs. We next attached the cystic EBs onto gelatin-coated plates and cultured for 5 days. In the results of real-time PCR and immunocytochemistry analysis, AFP-expressing cells migrated and localized at the outgrowth region of attached cystic EBs. To obtain the AFP-expressing cells of the outgrowth region, we manually isolated by using micro- dissection and cultured them. These cells strongly express AFP over 70% of isolated cells post re-plating. Here, we first showed an expression pattern of specifically localized ELCs by Activin-A during differentiation of hESCs. From this observation, we could highly purified ELCs from undifferentiated hESCs. Taken together, our system will provide a novel and efficient option to generate ELCs from hESCs.

The origin of squamous cell components in salivary gland tumor has been not yet clarified in detail. The squamous cell differentiation from adenocarcinoma has been reported in various carcinoma by HPV transfection in vitro. The adenocarcinoma cells adjacent to the squamous cell carcinoma components were positive for HPV. This is thought to indicate that after adenocarcinoma cells are transfected with HPV, they undergo morphological changes, and that squamous cell differentiation follows. The purpose of this study were to examine the effects of HPV-16 E6/E7 gene transfection into SGT cell line from human salivary gland adenocarcinoma, and to study the relation between the E6/E7 gene and squamous differentiation. Plasmid pBR322 containing HPV-16 was transfected into cultured SGT cell line using lipofectin method. Hygromycin was used as a selection marker. The presence of HPV E6/E7, transglutaminase 1, and involucrin mRNAs and protein in E6/E7 gene transfected cells was investigated by RT-PCR and immunoslot blot method. The apoptosis index was analysed by flow cytometry. The growth rate of E6/E7 gene transfected cells was reduced. E6/E7 transfected SGT cells increased apoptosis index. Involucrin and TGase I mRNAs by the squamous cell differentiation was most conspicuous in the E6/E7 gene transfected cell compared with non transfected cells. Squamous cell differentiation demonstrated in the transfectedSGT cell line, which expressed E6/E7 fusion gene mRNA.E6/E7 gene transfected cells showed squamous cell differentiation, expressing involucrin and TGase 1 protein by immunoslot blotting. The transfected SGT cell which expressed E6/E7 gene mRNA showed the squamous cell differentiation particularly clearly, and apoptosis was also demonstrated. It suggested that E6/E7 gene transfection into human salivary gland adenocarcinoma cells might induce clear squamous cell differentiation and contribute to study the pathogenesis of human salivary gland adenocarcinoma.

1999, 2002년에 인공교배한 조합에서 선발된 계통들을 2004년부터 2006년까지 특성검정을 거쳐 최종 육성한 분화장미 신품종 ‘타이니볼’은 백/분홍 복합색의 왜성형 분화장미로 대비품종에 비해 꽃크기는 작은 편이나 꽃 잎수와 착화수가 많으며, 수고가 낮아 왜화제 처리 없 이도 상품화율이 높다.

Mesenchymal stem cells (MSCs) have the multipotent capacity and this potential can be applied for obtaining valuable cell types which can use for cell therapy on various regenerative diseases. However, insufficient availability of cellular source is the major problem in cell therapy field using adult stem cell sources. Recently, human embryonic stem cells (hESCs) have been highlighted to overcome a limitation of adult cellular sources because they retain unlimited proliferation capacity and pluripotency. To use of hESCs in cell therapy, above all, animal pathogen free culture system and purification of a specific target cell population to avoid teratoma formation are required. In this study, we describe the differentiation of a mesenchymal stem cell-like cells population from feeder-free cultured hESCs(hESC-MSCs) using direct induction system. hESC-MSCs revealed characteristics similar to MSCs derived from bone marrow, and undifferentiated cell markers were extremely low in hESC-MSCs in RT- PCR, immunostaining and FACS analyses. Thus, this study proffer a basis of effective generation of specialized human mesenchymal stem cell types which can use for further clinical applications, from xenofree cultured hESCs using direct induction system.

배지의 적정 수분함량과 기상율은 분화식물 생산에 있어 중요한 환경 요인들이다. 매트관수시스템에서 Ardisia 소형분화 생산에 적합한 배지의 물리성을 구명 하고자 실험을 수행하였다. 실험에 사용된 공시 작물은 산호수와 자금우였다. 피트모스 배지에 왕겨와 펄라이트를 각각 부피비로 20, 40, 60%로 혼합하였다. 배지의 총공극은 왕겨의 혼합비율이 증가함에 따라 증가하였으나, 펄라이트는 혼합비율이 증가함에 따라 감소하였으며, 왕겨와 펄라이트의 혼합비율이 증가함에 따라 기상율은 증가하였으나, 배지의 수분함량은 감소하였다. 배지의 기상율은 왕겨 혼합배지에서 펄라이트 혼합배지보다 높았으며 증가율은 왕겨혼합배지에서 더 높았다. 배지의 CO2 농도는 기상율이 증가함에 따라 감소 하였는데, 왕겨 혼합배지에서 펄라이트 혼합배지보다 CO2 농도가 더 높았다. 산호수와 자금우의 생체중과 건물중은 왕겨를 60% 혼합한 배지에서 가장 높았으나, 지상부와 지하부의 건물 비율은 가장 낮았다. 매트관수 시스템에서 Ardisia속 식물인 산호수와 자금우의 소형 분화 생산에 적합한 배지의 물리성은 총공극과 기상율, 포트용수량이 각각 82.8, 25.6, 57.2%이었다.

It is not yet clear to know how normal human osteoblasts(NHost) from oral and maxillofacial area deposit, stabilize, and configure their extracellular matrix on dental biomaterial surfaces. Therefore it is necessary to design biomaterials with improved biocompatibility that will promote earlier bone differentiation and mineralization. There is now increasing evidence that TGase 2 may play a role in the initiation and regulation of the mineralization processes and serves to cross-link osteocalcin and osteopontin, which are predominant substrates for TGase 2 expressed during bone mineralization. But it is still unclear as to which TGase 2 is expressed by NHost in vitro bone formation. The purpose of this s tudy was t o determine the level of TGase 2 expression associated with collagen I , osteopontin and osteocalcin in NHost cell lines from oral and maxillofacial area in vitro. We will investigate whether TGase 2 has an active role in the biocompatibility of dental biomaterials during differentiation and mineralization of NHost. NHost cell lines were cultured under DMEM with 10% FBS at 37ﾟC and 5% CO2. Collagen quantification, mineralization and calcium assay, ALP activity assay, and RT-PCR analysis during bone differentiation and mineralization were done. ALP levels showed higher activity under AA+hGP t han under AA. I nhibition o f T Gase 2 by cystamine showed d ecreased Ca++ concentration, c ollagen I deposition and ALP level during 11 days of differentiation. TGase 2 mRNA expression of NHost was constant during mineralization stage. TGase 2 enzyme activity was increased during differentiation. Osteopontin mRNA expression during mineralization was higher than that of osteocalcin and collagen I . It suggested that TGase 2 associated with collagen I, osteocalcin and osteonectin might play a role in the differentiation of NHost from oral and maxillofacial area, but a little involved in mineralization.

Cementum is a hard connective tissue, produced by cementoblasts during tooth root formation, which provides for the attachment of the periodontal ligament to the roots and surrounding alveolar bone. Establishment of this attachment is an important event in the regeneration of lost periodontal tissues. We examined whether or not odontoblast conditioned media(CM) have a regulatory influence on the differentiation and mineralization of cementoblasts(murine cementoblastic cell line, OCCM-30) in vitro. To identify the effect of odontoblast conditioned media and dentin non collagenous proteins (dNCPs) on cementoblast differentiation and mineralization, we treated CM and dNCPs to cementoblast then differentiated the cells for 14 days. To evaluate the formation of mineralized nodules alizarin-red S staining was performed at 0,4,7 and 14 days. Expression of cementum matrix genes was measured by RT-PCR. Mineralization of cementoblasts was accelerated with CM from odontoblastic MDPC-23 and OD-11. The expression of BSP, ALP, and OC mRNA in cementoblastic OCCM-30 cells was facilitated by the MDPC-23 and OD-11 cells. The extracted dNCPs had little influence on the proliferation, cell cycle modification, and chemotaxis of OCCM-30 cells. Although the dNCPs did not exhibit chemotactic activities for cementoblasts, the dNCPs promoted the differentiation and mineralization of cementoblasts. In conclusion, the dentin matrix protein, or the secreted products of odontoblast, facilitates cementoblast differentiation and mineralization. This represents a new approach and suggests another avenue for cementum regeneration.

This study aims to identify the origin and usage of ‘-s n’, a suffix for male title, shown in dialects of Jeollanamdo, and based on the results, to describe the aspect of differentiation of Jeonnam dialects through the suffix of ‘-s n’. This study examines the different types of ‘-s n’, its usage, the meaning in term of dialect geography and characteristic of its social and the results are summarized as follows: First, previous studies sought for the origin of ‘-s n’ from ‘S nnim’, but this study is to find its origin from grammaticalization based on the dialect type of ‘S ngwon’ like ‘S won～Sewan’. Second, the title of ‘-s n’ has diverse sub-categories such as ‘name＋ last name＋s n’, ‘name of the place wives come from＋last name＋s n’, ‘name of previous residence＋last name＋s n’, ‘occupation＋last name＋ s n’, and ‘nickname＋last name＋s n’ in addition to ‘last name+s n’. However, according to social and cultural features of the community the title is used, especially, whether the community is nobility village (Banchon) or civilian village, its specific usage is different. Third, as the title of ‘-s n’ exist only in the eastern part of Jeonnam, it has different aspects from that of western part. In particular, it was connected with ‘-t' k’, a suffix of female title, and the identification following one’s husband was established instead of kinship terms, which works as an important factor that made Jeonnam dialects categorized into east and west. Fourth, today, the use of the title ‘-s n’ is limited to older people, in the civilian village rather than nobility village and used by men rather than women in higher rate.

Spring phenology of the oriental fruit moth, Grapholita molesta, was monitored using sex pheromone traps in apple cultivating areas. Their occurrence was earlier in southern areas and their population sizes were significantly different among orchards even in a local cultivating zone. The overwintering populations appeared to move between local orchards, based on the fact that monitoring data obtained at the sites between orchards were similar to those of nearby orchards. However, within orchards, these adult movements appeared to decrease and showed skewed occurrences at the side of upwind direction or close to neighboring orchards. At initial occurrence peak (April 20-25), the overwintering populations of the different localities were collected and analyzed in their genetic distances. PCR-RAPD analysis indicated that there were significant genetic differences among the overwintering populations of G. molesta. This genetic differentiation of overwinterin populations may be due to genetic bottleneck following differential selection pressures against the subpopulations of G. molesta during winter on the basis of the RAPD analysis that each early spring population was significantly different to its previous fall population in the same locality.

호접란 촉성재배를 위한 하계 고온기 폐광 자연냉풍 처리가 개화에 미치는 영향을 밝히고자 본 실험을 실 시하여 다음과 같은 결과를 얻었다. 보령지방 폐광의 자연냉풍온도는 12~14oC이였으며 이 온도를 주온 22oC, 야온 20oC로 조절하여 6월부터 8월까지 처리하 였다. 이 온도는 호접란 화성유도 기본 온도범위이었으 며, 이 때 외부 평균기온은 28.4~32.8oC로 나타났다. 화경 발생은 20일저온경과에서 3% 발생되었으며, 45일 경과에서 65% 이상 발생되었고, 90일 경과에서 100% 발생하였다. 화경장은 30일 경과에서 48.7cm로 짧았으며, 60일 경과에서 62.4cm로 길어져 처리기간이 길수록 화경장이 신장되었다. 화수장, 소화수도 같은 경향이었고, 화경 마디수는 6~7개로 저온 경과일수와 는 큰 영향이 없는 것으로 나타났다. 개화는 45일 경 과에서 9월 15일에 가장 빨리 개화되었고 저온 경과 일수가 길수록 늦어지는 경향을 나타내었다. 고온기 폐 광 자연냉풍처리는 무처리에 비하여 화경 발생시기는 3개월, 개화기는 4개월 정도 촉진되었다.

본 연구는 돼지 골수에서 존재하는 조혈 줄기 세포 및 전구 세포를 이용해 이종 동물 모델인 태아 마우스 복강 생체 이식을 통하여 돼지 조혈 세포의 이종 조혈 조직에서의 증식과 분화 특성을 규명하였다. 선천성 면역 부전 마우스인 NOD/SCID 마우스 태아 조혈 환경에 돼지 골수 유래 조혈 줄기 세포 및 전구 세포를 이식하고, 이식 후 5주령에 마우스 조혈기관에서의 돼지 조혈 세포의 증식과 분화 특성을 돼지 특이적 항체 면역 염색으로 유세포 분석을 실시한 결과, 마우스 조혈 조직인 골수, 흉선, 간장, 비장 및 림파절에서 돼지 조혈 세포의 분화 및 증식이 관찰되었다. 특히 돼지의 T 면역세포가 골수계 세포에 비해서 높은 chimerism이 관찰되어 태생 초기의 NOD/SCID 조혈 환경에 의한 특이적 T 면역세포의 증식에 적합한 조혈 환경을 제공하고 있다는 사실이 밝혀졌다. 본 마우스 신생 NOSD/SCID 복강 이식 동물 모델을 이용해 돼지 T 면역세포의 분화 발달 연구 및 이종 장기 이식 기전 연구에 좋은 모델로서 활용이 기대된다.

커피나무를 실내분화로 개발할 목적으로 광도에 따 른 생육반응과 겨울철 온도관리에 대한 실험을 수행한 결과는 다음과 같다. 50% 차광망으로 0~3겹까지 차광 하여 생육을 조사한 결과, 1겹과 2겹에서 생육이 가장 양호하고 엽색도 진한 녹색이며 잎에 광택이 있어 관 상가치도 가장 높은 것으로 나타났다. 3겹 차광에서는 엽색은 진하나, 생육이 지나치게 억제된 결과를 보였으 며, 무차광에서는 엽색이 탈색되고 생육도 현저하게 저 하되었다. 겨울철에 최저 온도를 5, 10, 15, 20oC로 설정하고 커피나무를 관리한 결과, 15oC 이상에서 정 상적인 생육이 가능했으며, 엽색도 진한 녹색으로 관상 가치가 높게 나타났다. 그러나 10oC에서는 생육이 현 저하게 저하되었고, 5oC에서는 생육저하뿐 아니라 일 부 개체는 저온피해로 고사되었다. 따라서 커피나무를 분화로 재배하고자 할 때는 50% 차광망으로 2겹이 가장 좋으며, 겨울철 야간 최저 온도는 15oC이라고 판단되었다.