• 한국습지학회지
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• 제2권2호
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• pp.117-131
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• 2000

강원도 인제군에 위치한 대암산 용늪(1,280m)의 습원 식물상과 식생에 대하여 Braun-Blanquet(1964) 의 방법으로 1998년 6월 13일부터 14일까지와 8월 7일부터 8일까지의 기간에 실시하였다. 습원내의 소산식물은 선태류 4과 4속 7종, 유관속식물 49과 127속 153종 30변종 1품종으로 총 191종류였다. 식생은 가는오이풀 군락군(Sanguisorba tenuifolia var. alba community group)으로 11개군락으로 구분되었다. 즉, 습원의 주요 식물군락은 삿갓 사초군락(Carex dispalata community), 물이끼 군락(Sphagnum palustre community)이었으며, 그 훼손지의 재생지역에는 솔잎사초 군락(Carex biwensis community), 청비녀골 풀 군락(Juncus papillosus community)이 함께하고 있었다. 또한 지하수위가 저하된 건조지역에는 진퍼리새 군락(Malinia japonica community)을 중심으로, 꼬리조팝나무 군락(Spiraea salicifolia community), 산새풀-꽃쥐손이 군락(Geranium eriosteman var. megalanthum-Calamagrostis langsdorffii community) 외에 골풀 군락(Juncus effusus var. decipiens community), 버드나무 군락(Salix koreensis community), 동의 나물 군락(Caltha palustris var. mem branacea community), 흑삼릉 군락(Sparganium stdoniferum community)으로 이루어져 있었다. 특히 그간에 실시되어온 1969년, 1976년, 1988년의 조사와 금번의 조사를 비교 분석해본 결과, 습원의 삿갓사초 군락, 물이끼 군락의 규모가 감소되고, 진퍼리새 군락이 침입되어 번식하며 그 규모가 커지고 있었다. 이는 스케이트장 조성과 가뭄의 영향 등 습원의 건조화 때문이나 인간의 출입과 훼손에 의한 교란이 없다면 고층습원의 회복 및 재생이 가능할 것으로 판단되었다.

• Journal of Ginseng Research
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• 제33권4호
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• pp.330-336
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• 2009

G-Rb1에 의한 단핵구 세포주인 U937 세포와 대식세포주인 RAW264.7 세포의 유착조절 능, 세포표면 단백질 발현율 및 세포모양 변화능 조절효과를 조사하여 다음과 같은 결론을 얻었다. 1. G-Rb1은 CD29항체 (MEM101A) 및 CD43 항체(161-46) 처리에 의해 유도된 세포-세포간 유착현상을 20%에서 35%까지 유의적으로 상승하였다. 2. G-Rb1은 fibronectin처리에 의해 유도된 U937 세포-fibronectin간 유착현상을 상승시켰다. 3. G-Rb1은 CD29 및 CD43의 세포표면 발현 수준을 유의적으로 증가시켰으나 CD82의 발현은 억제시켰다. 4. G-Rb1은 PMA에 의해 유도된 형태변화는 촉진경향을 반면에 LPS에 의한 변화는 억제경향을 나타내었다. 인삼의 주요사포닌 성분인 G-Rb1의 약리작용이 활발히 연구되고 있다. 특별히, 본 화합물이 갖는 면역증강 및 항암 효능은 인삼이 갖는 이들 효과를 대신하는 것으로 보고 되어져 있다. 그러나, 세포유착과정이 다양한 염증과정, 알러지 반응 및 암세포의 이동과 전이성 등에 관련이 있다는 관점에서 볼 때 본 연구결과는 이들 약물이 갖는 치료기전을 설명하기에는 충분하지 않는 것으로 판단된다. 오히려, 본 결과는 G-Rb1이 특정 농도 ($25-50\;{\mu}M$ 수준)에서 세포의 활성을 촉진할 수 있다는 가능성을 시사한다고 하겠다. 특별히, G-Rb1의 세포유착 촉진 기능에 관한 기전 이해를 위해, 이후, 세포유착 유도 신호전달 단백질의 활성을 포함한 다양한 분자적 수준에서의 추가적인 실험들을 진행하고 한다.

• 한국식품영양과학회지
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• 제31권2호
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• pp.236-240
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• 2002

본 연구에서는 타조 사육농장으로부터 폐기물로 나오는 타조알 껍질을 부가가치가 높은 칼슘자원으로 활용하고자, 타조알 껍질의 성분특성 및 젖산칼슘 제조를 위한 최적 회화 조건을 검토하였다. 타조알 껍질(ostrich egg shell with mem-brace, OESM)의 평균 무게는 255.17 g, 둘레는 39.50 cm, 길이는 15.20 cm 이었으며, 피막이 제거된 타조알 껍질(ostrich egg shell without membrane, OES)의 수분함량은 0.35%, 주요 무기질 성분은 Ca으로 전체의 40.98%를 차지하였으며, 단백질 함량은 2.43%, 아미노산 함량은 235.0 mg/100g을 나타내었다. OES로부터 회화분을 제조시, 회화 최적시간은 $700^{\circ}C$에서는 12시간, 80$0^{\circ}C$에서 는 80분, 90$0^{\circ}C$에서는 15분이었으며, 이때 수율은 54.5~54.6%, $L^{*}$ 값은 97.26~97.51, $a^{*}$ 값은 -0.30~-0.34, $b^{*}$ 값은 0.63~0.98 범주를 나타내었다. OES의 입자를 0.841 mm이상, 0.425~0.250mm사이, 0.150 mm 이하의 크기로 구분하여 90$0^{\circ}C$에서 15분간 회화한 후 그 수율(%)을 비교한 결과, 입자 크지에 따른 회화분의 수율에는 유의적인 차이가 없었다.

• Journal of Acupuncture Research
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• 제23권5호
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• pp.69-77
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• 2006

Background : Mesenchymal stem cells (MSCs) are present in most of the tissue matrix, taking part in their regeneration when injury or damage occurs. The aim of this study was to investigate the presence of cells with pluripotential characteristics in human subchondral bone and the capacity of these cells to differentiate to osteoblast. Methods : Human subchondral bone were digested with collagenase. Isolated cells were cultured with a-MEM, 15% FBS, 10-8M dexamethasone and 50 ng/mL ascoric acid. Cells from 0 day(isolated cells), 7 day (first subculture) and 14 days (third subculture) were used to carry out phenotypic characterization experiments flowcytometry analysis with 11 monoclonal antibodies) and osteogenic differentiation experiments. Osteogenic differentiation of cells was assessment by quantification of bone extracellular matrix components by following analysis: alkaline phosphatase(ALP) stains to detect ALP activity, RT-PCR and western blot to detect osteocalcin (OCN), osteopontin (OPN) and type I collagen(Col I), and Alizarin red stains to detect calcium deposition. Results : Flowcytometry analyses showed that in our population more than 98% of cells were positive for MSC markers: SH-2(CD105, 99%), CD29 (95%), CD73 (95%). Cells were negative for hematopoietic markers (CD11b, CD34, and CD45). Furthermore, cells showed positive stain to multipotent markers such as CDl17 (c-kit) (15.1%), and CD166 (74.9%), and cell adhesion molecules such as CD54 (78.1%) and CD106 (63.5%). The osteogenic specific marker analyses showed that the culture of these cells for 7 and 14 days stimulates ALP, OCN, OPN and Col I synthesis by RT-PCR and Western blot analysis. Also, after 14 days in the culture of MSCs induces mineralization by Arizarin red stain. Conclusion : In this work, we demonstrated a new and efficient method for osteoblastic differentiation of human subchondral bone stem cells. As MSCs takes part in reparative processes of adult tissues, these cells could play an important role in osteogenesis.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제31권6호
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• pp.461-467
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• 2005

Background. 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, succinimidyl ester mixed (CFSE) is the fluorescent labelling agent of living cells and used to trace the cells in vivo after transplatnation of various cells. The CFSE labelled cells can maintain fluorescence for up to 7 days after labelling. The MC3T3-E1 cell line (MC3T3) has been used for many studies about osteoblast, which is well known as a mouse preosteoblast. So the CFSE would be used to trace the transplanted MC3T3. However there are few reports about CFSE labelling of MC3T3. This study is aimed to know about adequate concenturation and incubation time of CFSE to MC3T3. Materials and methods. The MC3T3 was incubated in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ using ${\alpha}$-minimal essential medium (${alpha}$-MEM) containing10% FBS and gentamycin. Ten mM CFSE solution in dimethylsulphoxide (DMSO: 1%) was diluted with phosphate buffered saline (PBS) and final concentration of culture medium was, respectively, 5, 10, 15, 20, 25 and 30 ${{\mu}M$. Then the MC3T3 was incubated with CFSE in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 and 45 minutes in each concentration. The fluorescence of CFSE labelled cells was analysed with a inverted fluorescence microscope. The duration of cell labelling was also studied. Trypan blue dye exclusion test was done for cell viability. Results. For concentration between 5 and 10 ${\mu}M$, CFSE did not significantly label the MC3T3 in vitro. The destruction of MC3T3 was observed at the concentration of 20 ${\mu}M$. In the concentration of 15 ${\mu}M$, the best labelling was obtained at an incubation period between 15 and 30 minutes. The MC3T3 labelled with an incubation period of 15 minutes at 15 ${\mu}M$ was still fluorescent 7 days after CFSE labelling. The mean cell viability was 95.93%. Conclusion. These results suggests an incubation period of 15 minutes at 15 ${\mu}M$ of CFSE provides best labelling of MC3T3 in vitro.

• Tuberculosis and Respiratory Diseases
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• 제69권6호
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• pp.456-464
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• 2010

Background: Synergistic antitumor effects of the combined chemoimmunotherapy based on dendritic cells have been reported recently. The aim of this study is to search new applicability of gefitinib into the combination treatment through the confirmation of gefitinib effects on the monocyte derived dendritic cells (moDCs); most potent antigen presenting cell (APC). Methods: Immature and mature monocyte-derived dendritic cell (im, mMoDC)s were generated from peripheral blood monocyte (PBMC) in Opti-MEM culture medium supplemented with IL-4, GM-CSF and cocktail, consisting of TNF-${\alpha}$ (10 ng/mL), IL-$1{\beta}$ (10 ng/mL), IL-6 (1,000 U/mL) and $PGE_2$ ($1{\mu}/mL$). Various concentrations of gefitinib also added on day 6 to see the influence on immature and mature MoDCs. Immunophenotyping of DCs under the gefitinib was performed by using monoclonal antibodies (CD14, CD80, CD83, CD86, HLA-ABC, HLA-DR). Supernatant IL-12 production and apoptosis of DCs was evaluated. And MLR assay with $[^3H]$-thymidine uptake assay was done. Results: Expression of CD83, MHC I were decreased in mMoDCs and MHC I was decreased in imMoDCs under gefitinib. IL-12 production from mMoDCs was decreased under $10{\mu}M$ of gefitinib sinificantly. Differences of T cell proliferation capacity were not observed in each concentration of geftinib. Conclusion: In spite of decreased expressions of some dendritic cell surface molecules and IL-12 production under $10{\mu}M$ of gefitinib, significant negative influences of gefitinib in antigen presenting capacity and T cell stimulation were not observed.

• Clinical and Experimental Reproductive Medicine
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• 제32권3호
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• pp.207-216
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• 2005

Objective: To understand the crucial requirement for the normal early folliculogenesis, we evaluated molecular as well as physiological differences during in vitro ovarian culture. Among the important regulators for follicle development, anti-Müllerian hormone (AMH) and FSH Receptor (FSHR) have been known to be expressed in the cuboidal granulosa cells. Meanwhile, it is known that c-kit is germ cell-specific and GDF-9 is also oocyte-specific regulator. To evaluate the functional requirement for the competence of normal follicular development, we investigated the differential mRNA expression of several factors secreted from granulosa cells and oocytes between in vivo and in vitro developed ovaries. Materials and Methods: Ovaries from ICR neonates (the day of birth) were cultured for 4 days (for primordial to primary transition) or 8 days (for secondary follicle formation) in ${\alpha}$-MEM glutamax supplemented with 3 mg/ml BSA without serum or growth factors. The mRNA levels of the several factors were investigated by quantitative real-time PCR analysis. Freshly isolated 0-, 4-, and 8-day-old ovaries were used as control. Results: The mRNA of AMH and FSHR as granulosa cell factors was highly increased according to the ovarian development in both of 4- and 8-day-old control. However, the mRNA expression was not induced in both of 4- and 8-day in vitro cultured ovaries. The mRNA expression of GDF-9 known to regulate follicle growth as an oocyte factor was different between in vivo and in vitro developed ovaries. In addition, the transcript of GDF-9 was expressed in the primordial follicles of mouse ovaries. The mRNA expression of c-kit was not significantly different during the early folliculogenesis in vitro. Conclusion: This is the first report regarding endogenous AMH and FSHR expression during the early folliculogenesis in vitro. In conclusion, it will be very valuable to evaluate cuboidal granulosa cell factors as functional marker(s) for normal early folliculogenesis in vitro.

• Clinical and Experimental Reproductive Medicine
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• 제24권2호
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• pp.241-251
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• 1997

체외성숙과 수정된 돼지 난자의 체외발달능이 체외배발생 배양액인 NCSU 배양액에 0.4% BSA, 10% 혈청 혹은 아미노산 (2% BME 아미노산 용액과 1% MEM 아미노산 용액)을 첨가함으로서 조사되었다. 본 실험에 공시된 난자는 체외수정 추 30시간 (2-세포기)혹은 48 시간 ($2{\sim}4$-세포기)에 회수하였다. 실험I에서 0.4% BSA가 첨가된 NCSU 배양액에서 2-세포기 난자들의 배양경과시간에 따른 발달능을 조사한 결과, 배양 후 72 시간 (체외수정 후 102 시간)에 상실배기와 배반포기 배가 나타났으며, 배양 후 120 시간째 (체외수정 후 150 시간)에도 팽창된 배반포기 배까지만 발달하였다. 실험II는 체외수정 후 48 시간의 분할된 ($2{\sim}8$-세포기) 난자들의 핵과 외관적 분할구와의 수적 차이를 조사한 결과, $2{\sim}4$-세포기보다는 5-세포기 이상에서 핵과 분할구의 조화에 차이가 많았다. 실험III에서는 $2{\sim}4$-세포기 난자들을 배양후 5일째의 배반포들의 투명대의 두께, 난자 크기 그리고 inner cell mass (ICM)과 trophectoderm (TE)의 세포 배열을 조사한 결과, 난자의 크기가 커짐에 따라서 투명대가 얇아지고 전체 세포수가 증가하였지만, ICM의 비율은 차이가 없었다. 실험IV에서는 BSA, 혈청 혹은 아미노산이 첨가 혹은 무첨가된 배양액내에서 $2{\sim}4$-세포기 난자들의 배반포 후 부화능력을 조사한 결과, 모든 군에 있는 난자들은 팽창된 배반포기 배까지 발달할 수 있었던 반면, 난자의 부화는 아미노산 혹은 혈청이 포함된 배양액에서만 일어났다. 더우기 상실배기와 배반포기 시기에 혈청의 첨가는 부화 배반포기 배의 발달을 현저히 증가시켰다. 또한 아미노산과 혈청의 영향을 받은 팽창 배반포기 배는 얇은 투명대, 팽창된 난자의 크기 그리고 ICM과 전체 세포수의 증가를 보였다. 이상의 결과로 미루어 볼때, 배양액내에 대한 아미노산과 혈청의 첨가는 돼지 배반포기 배의 부화를 유도할 수 있다고 보며, 더우기 이들 요소들은 투명대의 두께, 난자의 크기 그리고 ICM과 전체 세포수에 영향을 미친다.

• 대한구강악안면병리학회지
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• 제42권5호
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• pp.111-118
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• 2018

Nicotine of tobacco component has a controversial impact in the clinical outcome of dental implants. Although numerous nicotine effects on bone healing around implants have been presented, it is rarely reported in vitro study about normal human osteoblast(NHost) from oral and maxillofacial area at the surface of implants. The purpose of the present study was to evaluate the effect of nicotine on the proliferation and differentiation response of NHost to plasmatic and salivary levels of nicotine reported in smokers at the surface of screw-type plasma-sprayed titanium implants. NHosts were seeded on the surface of titanium implants and cultured for 21 days in ${\alpha}-MEM$ supplemented with 10% FBS, 50mg/ml ascorbic acid, 5mM ${\beta}$-glycerophosphate and 100nM dexamethasone. Seeded implants were exposed to various nicotine concentration(0.05-0.5mg/ml) from 1 to 21 days, and characterized for cell morphology, proliferation, differentiation, alkaline phosphatase(ALP) activity and ionized calcium concentration(Cai) of medium. Continuous exposure to higher nicotine concentration(above 0.3mg/ml) induced a dose- and time-dependent vacuolation of the cytoplasm, and a tendency to detach from the implant surface. 0.05mg/ml(lower nicotine concentration) did not cause significant effects in the cell proliferation and ALP activity. 0.1-0.2mg/ml caused evident dose-dependent effects in increased cell proliferation, ALP activity and earlier onset of matrix mineralization at levels up to 0.2mg/ml, while a dose-dependent inhibitory effect at 0.3-0.5mg/ml. Cai concentration of control group was decreased at 14 days. Increased Cai concentration at 0.1-0.2mg/ml, decreased Cai concentration at 0.3mg/ml and no change at 0.5mg/ml during the culture period were seen. It suggested that nicotine concentration could paly an role in modulating NHost activity as a contributing factor associated with proliferation and differentiation of NHost at the surface of implants.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.30-30
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• 2001

The culture of preantral follicles has important biotechnological implications through its potential to produce the large quantity of oocytes for embryo production, transgenesis research, conservation of rare breed, and a potential source of ovarian genetic material. The present study was conducted to establish the optimal conditions of in vitro culture for intact bovine preantral follicles; and to examine the developmental ability of oocytes derived from the in vitro-grown preantral follicles; and to investigate the effects of various concentrations of FSH and LH on these processes. Bovine preantral follicles (150 $\pm$ 1.2${\mu}{\textrm}{m}$), surrounded by theca cell, were isolated enzymetically and mechanically from ovarian cortical slides in Leibovitz L-15 medium containing 1 mg/$m\ell$ collagens and 0.2 mg/$m\ell$ DNase I and cultured for 25 days in the presence of different concentrations of bovine FSH and LH in $\alpha$MEM medium with insulin, transferrin, and selenite. The survival was tested by frypan Blue and Hematoxylin. The survival and growth rates of follicles were higher in FSH treatment groups than these in control (P<0.001), but there were no significant differences between the LH treatment groups and the control. In 25 days, the survival and growth rates of follicles in FSH and LH treatment group (50%, 300$\pm$1.0${\mu}{\textrm}{m}$) were higher than in FSH treatment group (40%, 244$\pm$0.5${\mu}{\textrm}{m}$) and the control group (25%, 160$\pm$ 1.0${\mu}{\textrm}{m}$). Fifty-five percent of healthy antral follicles were obtained, and 60% of the oocytes complete meiotic maturation to the metaphase II stage. Twenty-two percent of the mature oocytes underwent cleavage, and 9% developed to the blastocyst stage. In this study, in vitro-grown oocytes (111 $\pm$ $1.5mutextrm{m}$), under our culture conditions, were not equivalent in size to the in vivo-grown oocytes (130$\pm$1.3${\mu}{\textrm}{m}$). Therefore, these results suggest that bovine preantral follicles with intact theca cell can grow to the antral stage in 25days, and that oocytes from those follicles can acquire the meiotic competence and normally undergo fertilization and development to the blastocyst stage. However, the developmental capacity of in vitro-grown oocytes is presumably not comparable to those of the in vivo counterparts.