• Title/Summary/Keyword: 줄기 증식

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Effect of Ursolic Acid on the Development of Mouse Embryonic Stem Cells under Hypoxia (저산소 상태에서 우르솔산이 배아줄기세포 성장에 미치는 효과)

  • Han, Gi Yeon;Park, Jae Hong;Oh, Keon Bong;Lee, Sei-Jung
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1223-1229
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    • 2013
  • Ursolic acid (UA) a bio-active ingredient found in a variety of fruits and vegetables, and it has potent antioxidant activity. However, the role of UA in mouse embryonic stem (ES) cells is poorly understood. This study investigated the functional role of UA in regulating the development of mouse ES cells under hypoxia. Hypoxia did not exert a significant effect on the undifferentiated state of mouse ES cells. However, it induced reactive oxygen species (ROS) generation and increased the level of lactate dehydrogenase (LDH) production at 48 h of hypoxic exposure. Conversely, oxidative stress induced by hypoxia was significantly inhibited by UA ($30{\mu}M$) pretreatment. Hypoxia significantly decreased cell survival and the level of [$^3H$] thymidine incorporation, both of which recovered following pretreatment of UA. In addition, UA decreased the apoptotic effect of hypoxia by attenuating caspase-3 cleavage or by recovering cellular inhibition of the apoptotic protein (cIAP)-2 and Bcl-2 expression. We further found that UA decreased senescence-associated beta-galactosidase activity. We suggest that UA is a natural antioxidant and one of the functional modulators of hypoxia-induced survival, apoptosis, proliferation, and aging in mouse ES cells.

Role of NFAT5 in Osteogenic Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells (인체 지방 유래 중간엽 줄기세포의 골분화 조절 기전에서 NFAT5의 역할)

  • Lee, Sun Young;Yang, Ji won;Jung, Jin Sup
    • Journal of Life Science
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    • v.23 no.4
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    • pp.471-478
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    • 2013
  • Human adipose tissue-derived mesenchymal stem cells (hADSCs) have therapeutic potential, including the ability to self-renew and differentiate into multiple lineages. Understanding of molecular mechanisms of stem cell differentiation is important for improving the therapeutic efficacies of stem cell transplantation. In this study, we determined the role of nuclear factor of activated T cells (NFAT5) in the osteogenic differentiation of hADSCs. The down-regulation of NFAT5 expression by the transfection of a specific siRNA significantly inhibited osteogenic differentiation of hADSCs and decreased the activity of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$) promoter without affecting their proliferation and adipogenic differentiation. The inhibition of NFAT5 expression inhibited the basal and Tumor Necrosis Factor ${\alpha}$ (TNF-${\alpha}$) induced activation of NF-${\kappa}B$, but it did not affect TNF-${\alpha}$-induced degradation of the $I{\kappa}B$ protein. These findings indicate that NFAT5 plays an important role in the osteogenic differentiation of hADSCs through the modulation of the NF-${\kappa}B$ pathway.

Optimizing In Vitro Propagation of Sophora koreensis Nakai using Statistical Analysis (다양한 통계분석 기법을 이용한 개느삼(Sophora koreensis Nakai)의 기내 증식 최적 조건 구명)

  • Jeong, Ukhan;Lee, Hwa;Park, Sanghee;Cheong, Eun Ju
    • Journal of Korean Society of Forest Science
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    • v.110 no.1
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    • pp.53-63
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    • 2021
  • Sophora koreensis Nakai is an indigenous plant in Koreawith a restricted natural range, part of which is in Gangwon province. The species is known to contain phytochemicals that have beneficial effects on human health, and it is economically important in bioindustry. Because of the limited number of plants in a small range of habitats, the mass-propagation method should be developed for use and conservation. In vitro tissue culture is a reliable method in terms of mass propagation from selected clones of the species. We investigated the optimal conditions of the medium in this process, especially focusing on the concentrations of plant growth regulators(PGRs) in the culture of stem-containing axillary buds. Three statistical methods, i.e., ANOVA, response surface method(RSM), and fuzzy clustering were used to analyze the plant growth, number of shoots induced, and shoot length with various combinations of PGRs. Results from the RSM differed from those of the other two methods; thus, the method was not suitable. ANOVA and fuzzy clustering showed similar results. However, more accurate results were obtained using fuzzy clustering because it provided a probability for each treatment. On the basis of the fuzzy clustering analysis, stem tissue produced the greatest number of shoots(11.03 per explant; 63.33%) on a medium supplemented with 5-��M 6-benzylaminopurine and 2.5-��M thidiazuron(TDZ). Proliferation of shoots(2.18 ± 0.21 cm, 63.33%) was attained on a medium supplemented with 2.5-��M BA, 2.5-��M TDZ, and 2.5-��M gibberellic acid.

In Vitro Propagation Using Stool Shoots of Mature Betula platyphylla var. japonica (자작나무 성숙목의 근주맹아를 이용한 기내증식)

  • Moon, H.K.;Youn, Y.;Hyun, Y.I.;Lee, S.K.
    • Journal of Korean Society of Forest Science
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    • v.80 no.4
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    • pp.416-419
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    • 1991
  • Effective micropropagation was achieved by axillary bud culture from stool shoots of 15-year-old Betula platyphylla var. japonica. Shoot development and proliferation from the explants were successful on WPM supplemented with 0.5 or 1.0mg/l BAP. All the regenerated shoots rooted when transfered to GD medium containing 0.2mg/l IBA. After transplaning to soil more than 95% of the plantlets survived and showed normal growth. The results demonstrate that masspropagation of selected mature trees is feasible using tissue culture technique.

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Composition of a Medium for Serum-free Culture of an Adipose-derived Stem Cell Line Established with a Simian Virus 40 T Antigen (Simian virus 40의 T항원 도입으로 수립한 지방유래줄기세포주의 효율적인 무혈청 배양법 및 무혈청 배지조성)

  • Kim, Gyu Bin;Joo, Woo Hong;Kim, Dong Wan
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1301-1307
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    • 2014
  • Adipose-derived stem cells (ADSCs) are considered promising tools for tissue regeneration. However, ADSCs have very poor proliferation capacity. Therefore, fetal bovine serum (FBS) is generally added to the culture media of ADSCs. As FBS contains many uncharacterized components that may affect cellular functions, methods for serum-free cultures of ADSCs have been widely investigated. In this study, to develop an efficient method for a serum-free culture of ADSC-T, we used an ADSC line established by introducing the simian virus 40 (SV40) T gene into primary ADSCs. We then investigated the effect of amino acids, vitamins, and other components on the growth of ADSC-T. When the ADSC-T cells were plated with DMEM/F12 serum-free medium, the cells did not proliferate, and the mixture of amino acids, vitamins, and B27 supplement did not increase the growth of the cells. However, when the ADSC-T cells were provided with serum-free DMEM/F12 after they had been cultured with serum-supplemented DMEM for 24 h, the cells proliferated, and the vitamins and B27 supplement increased the cell growth. Stem-Pro serum-free medium also appeared to be useful as a suspension culture for the ADSC-T cells. The ADSC-T cells secreted large amounts of proteins of around 70 kDa. Insulin-like growth factor (IGF) and fibroblast growth factor basic (FGF basic) were secreted by ADSC-T in larger amounts in the serum-free culture than in the serum-supplemented culture.

골조직 재생 촉진을 위한 Novel Perfusion Bioreactor

  • Lee, Si-U;Heo, Su-Jin;Jang, Ji-Yeon;Jeong, Jae-Yeong;Kim, Seung-Eon;Sin, Jeong-Uk
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.05a
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    • pp.46.2-46.2
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    • 2009
  • In vitro 상에서 골조직을 원활하게 재생하기 위해서는 3차원 지지체를 이용한 세포 배양과 세포 배양 시 세포의 형태와 기능을 유지/향상시키기 위한 인체 내 미세 환경 재현은 필수적이다. 따라서 본 연구에서는 뼈 성분과 유사한 생체 활성 물질인 hydroxyapatite (HA)와 생분해성 고분자인 poly $\varepsilon$-caprolactone (PCL)를 복합재료로 이용하여 내부 연결성이 우수한 골조직 재생용 3차원 지지체를 제작하였으며, 골 재생 능력 향상을 위하여 인체내 골조직의 기계적 미세 환경을 체외에서 구현한 새로운 형태의 perfusion bioreactor system을 개발/적용하였다. 또한 본 연구에서 개발된 perfusion bioreactor system의 생물학적 평가를 위해 MG63 (osteoblast like cell, 한국 세포주 은행)과 New Zealand White Rabbit에서 분리한 중간엽 줄기세포를 골조직 재생용 3차원 지지체에 파종하였다. 48시간 동안 안정화 후 perfusion bioreactor system을 이용하여 기계적 자극을 파종된 세포에 인가하였으며, 배양 기간 동안 세포의 증식 확인 및 형태학적 관찰을 실시하였다. 본 연구 결과, perfusion bioreactor system을 이용하여 기계적 자극을 인가한 실험군에서 세포의 증식 및 활성도가 대조군에 비해 우수함을 확인 할 수 있었다. 따라서, perfusion bioreactor를 이용한 세포 배양은 세포의 활성 향상 및 골조직 재생에 도움이 될 것으로 사료된다. 차후 perfusion bioreactor를 이용한 다양한 패턴의 자극이 골재생 능력 및 중간엽 줄기세포의 골 분화능에 미치는 영향에 대한 연구가 필요할 것으로 사료된다.

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Differentiation and Proliferation of Porcine T Lymphocytes in NOD/SCID Mice (NOD/SCID 모델 마우스 생체 내 돼지 T 면역세포의 증식 및 분화)

  • Lee, Yong-Soo;Kim, Tae-Sik;Kim, Jae-Hwan;Chung, Hak-Jae;Park, Jin-Ki;Chang, Won-Kyong;Kim, Dong-Ku
    • Reproductive and Developmental Biology
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    • v.31 no.1
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    • pp.1-6
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    • 2007
  • The nonobese diabetic / severe combined immune deficiency (NOD/SCID) has been used for determination of proliferation and differentiation of hematopoietic stem cells as xenotransplantation animal model. In this study, we transplanted porcine hematopoietic cells from bone marrow into NOD/SCID mice via intravenous injection to confirm the activity of differentiation and proliferation for porcine hematopoietic cells in vivo. Interestingly, we observed the result of high efficiency with pig T lymphocytes in hematopoietic organs, liver, spleen lymph node, and bone marrow in NOD/SCID mice. The porcine $CD3^{+}$ T cells were detected with $5.4{\pm}1.9%$ in bone marrow, $15.4{\pm}7.3%$ in spleen, $21.3{\pm}1.4%$ in liver, and $33.5{\pm}32.8%$ in lymph node of NOD/SCID mice at 6 weeks after trans-plantation Furthermore, immunohistochemical analysis showed the high engraftment of porcine T lymphocytes in spleen of NOD/SCID mice. Our data suggest that NOD/SCID mice are excellent animal model to determinate the generation md function of pig T lymphocytes.

Clinical Application of Autologous Adipose Tissue Derived Mesenchymal Stem Cells in Five Dogs with Stifle Joint Osteoarthrosis (무릎 골관절증을 보이는 개에서 자가지방유래 중배엽성 줄기세포 치료 다섯 증례)

  • Yoon, Hun-Young;Kang, Dong Jun;Lee, Soo-Han;Jeong, Soon-Wuk;Chung, Byung-Hyun
    • Journal of Veterinary Clinics
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    • v.31 no.3
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    • pp.253-257
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    • 2014
  • Five dogs presented with a history of pelvic limb lameness. On physical examination of the stifle joints, five dogs had pain, lameness, patellar luxation, or ligamentous instability. Craniocaudal and mediolateral radiographic projections revealed osteophytes or subchondral cystic lesions on the stifle joints. Based on a previously described Osteoarthrosis (OA) scoring technique, five dogs showed high OA scores. Combination of surgery and implantation of autologous adipose tissue derived mesenchymal stem cells (aAT-MSCs) or percutaneous injection of aAT-MSCs was determined with informed consent. $1{\times}10^6$ aAT-MSCs suspended in PBS was injected in the stifle joints. The follow-ups were completed 12 months after surgery. The follow-up information was based on physical examination by veterinarians. The lameness, pain on manipulation, and OA scores improved six or 12 months after implantation of aAT-MSCs. There was a radiographic evidence of decreased osteophytes and subchondral cystic lesions. These results suggest that implantation of aAT-MSCs can be considered an option for management of cases of OA in the stifle joints.

Study on Practical Micropropagation of Juiube Cultivars through Axillary Bud Culture (액아배양(腋芽培養)에 의한 신품종(新品種) 대추나무의 실용적(實用的)인 대량증식(大量增殖)에 관(關)한 연구(硏究))

  • Kim, Deok Sik;Lee, Sei Pyo
    • Journal of Korean Society of Forest Science
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    • v.77 no.4
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    • pp.445-452
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    • 1988
  • This study was conducted to establish practical micropropagation of jujube cultivars ('Geumsumg', 'Bokjo') by axillary bud culture. The results are summerized as follows : 1. Addition of activated charcoal to half-strength Murashige and Skoog(MS) medium supplemented with 0.5mg/l benzylaminopurine(BAP) enhanced shoot and root growth. At 500mg/l activated charcoal level 'Geumsung' showed best result, and shoot length and the number of multiple shoot were 6.4cm and 10.0, respectively. At 1,000mg/l activated charcoal level 'Bokjo showed best result, and shoot length and the number of multiple shoot were 7.5cm and 12.4, respectively. 2. As indole-3-butyric acid(IBA) concentration increased, rooting and callus growth of microshoot were enhanced. The optimum IBA concentration for shoot elongation and multiplication was 1.0mg/l. 3. Growth responses of shoot-tip and axillary bud segments between two jujube cultivars were different. 'Geumsung' showed that axillary bud explants were about twice better than shoot-tip explants for shoot multiplication, but 'Bokjo' showed that shoot-tip explants mere better than axillary bud explants for shoot elongation and multiplication. 4. In acclimatization processes of plantlets produced in vitro, the survival of plantlets with only root primordia in soil medium was better than that of plantlets with several routs resulting in 97.8%. 5. In cutting of in vitro-derived microshoot, paclobutrazol was more effective than IBA, naphth-aleneacetic acid(NAA) and $Rooton^{(R)}$ in rooting and root growth.

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Effects of Nutrient Solution Composition and Cutting Size on Growth of Virus-free Sweet Potato Plant in Nutrient Film Technique (NFT 수경재배에서 양액 종류 및 삽수 크기가 고구마 바이러스 무병주 생육에 미치는 영향)

  • Yoo, Kyoung-Ran;Lee, Seung-Yeob;Bae, Jong-Hyang
    • Horticultural Science & Technology
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    • v.30 no.6
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    • pp.686-693
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    • 2012
  • To develop a technique for mass-propagation of virus-free sweet potato [Ipomoea batatas (L.) Lam.] plant using nutrient film technique (NFT), the growth characteristics of 4 cultivars as affected by nutrient solution composition and cutting size were investigated. 72 cells (35 mL/cell) plug trays filled with vermiculite and perlite (1:1, v/v) were used. Vine length, fresh and dry weights of virus-free plants were the greatest in the nutrient solution recommended by National Horticultural Research Station in Japan, followed by that recommended by National Institute of Horticultural & Herbal Science in Korea, and Yamazaki's nutrient solution for lettuce. The growth of uppershoot cuttings was the best among 4 subsections of cutting. Vine length, and fresh and dry weights increased in the longer cutting treatments, and were better in 'Shinzami' and 'Yeonhwangmi' than those in 'Mannami' and 'Shincheonmi'. Vine diameter and length of the longest root were not significantly affected by the cutting size and cutting source. The growth characteristics of the single node cutting were not significantly different from those in 2-node cutting. The efficiency of rapid mass-propagation could be promoted with single node cuttings and uppershoot cuttings grown in NFT system.