• Title/Summary/Keyword: stem model

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Ovarian cell aggregate culture in teleost, marine medaka (Oryzias dancena): basic culture conditions and characterization

  • Jae Hoon, Choi;Seung Pyo Gong
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.1
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    • pp.19-30
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    • 2024
  • Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

Establishment of Embryonic Stem Cell Line from Isolated Blastomeres from Mouse Preimplantation Embryos (생쥐 초기 배아에서 분리한 할구를 이용한 배아줄기세포주 확립)

  • Lim, Chun Kyu;Sung, Ji Hye;Choi, Hye Won;Cho, Jae Won;Shin, Mi Ra;Jun, Jin Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.1
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    • pp.25-33
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    • 2006
  • Objective: The aim of this study was to investigate whether embryonic stem (ES) cells can be established from isolated blastomeres of mouse embryos. Methods: Blastomeres were separated from mouse (C57Bl/6J) 2- or 4-cell embryos. Isolated blastomeres or whole 4-cell embryos were co-cultured with mitosis-arrested STO feeder cells in DMEM supplemented with recombinant murine leukemia inhibitory factor and ES-qualified fetal bovine serum. After the tentative ES cell lines were maintained from isolated blastomeres or whole embryos, some of them were frozen and the others were sub-cultured continually. Characteristics of tentative ES cell lines as were evaluated for specific genes expressions with immunocytochemistry and RT-PCR. Results: One ES cell line (3.0%) was established from isolated blastomere of 2-cell embryo and one cell line (4.0%) from isolated two blastomeres of 4-cell embryo. And five cell lines (16.7%) were established from whole 4-cell embryos. Both cell lines from isolated blastomere and whole embryo expressed mouse ES cell specific markers such as SSEA-1, Oct-4 and alkaline phosphatase. Marker genes of three germ layers were expressed from embryoid bodies of both cell lines. Conclusion: This study suggests that mouse ES cells could be established from isolated blastomeres, although the efficiency is lower than whole embryos. This animal model could be applied to establishment of autologous human ES cells from biopsied blastomeres of preimplantation embryos in human IVF-ET program.

PRELIMINARY STUDY OF MENTAL REPRESENTATIONS OF PRESCHOOL CHILDREN EXPERIENCING SINGLE, SEVERE TRAUMA (심한 정신적 외상 경험을 한 학령 전기 아동의 정신적 표상에 대한 예비연구)

  • Eon, So-Yong;Song, Won-Woung;Oh, Kyung-Ja;Choi, Eui-Gyum;Shim, Eun-Ji;Shin, Yee-Jin
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.15 no.1
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    • pp.61-74
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    • 2004
  • Objectives:This study was performed to introduce a psychoeducational family therapy model for the families of schizophrenic patient and to investigate the effect of this model on the changes in coping style and depressive symptoms of the family members, and in perception of emotional support by families and depressive symptoms of patients. Methods:Nine preschool children, 3-5 years old, experiencing physical injuries caused by attack from a psychotic patient at kindergarten, were evaluated for psychological assessments;Intelligence test, MSSB(MacArthur Story-Stem Battery), H-T-P test(House-Tree-Person test). And their parents completed rating scale, KPI-C(Korean Personality Inventory for Children about children’s psychological conditions). Results:With respects to the contents and emotional reactions of MSSB, 9 preschool children showed generally high levels of anxiety, depression, avoidance, aggression, probably related to the traumatic experiences. Even though children couldn't verbally report directly about their traumatic experiences, in both MSSB, structured play narrative assessment tool, and HPT, free drawing and association test, they demonstrated psychiatric problems through reenactment plays, regardless of clinical diagnoses. Conclusion:Present study allowed us the chance to see beyond the outer pathological behaviors of PTSD in preschool children, through deeper evaluations of their mental representation. These preliminary data suggest deep understanding of internal representation would be of help for thorough evaluations and treatment plan for preschool children, experiencing severe trauma.

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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.

Age and life history of an old black pine (Pinus thunbergii Parl.) tree at Cave Temple on Mt. Sanbangsan, Jeju Island, Korea, died due to pine wilt disease in 2013

  • Kim, Eun-Shik;Lee, So-Hee;Kim, Joon-Bum;Kim, Chan-Soo;Yoon, Bong-Taek;Lee, Sung-Hoon;Lim, Wontaek;Kim, Hyojung;Choi, Junghwan;Han, Hyerim
    • Journal of Ecology and Environment
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    • v.38 no.1
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    • pp.85-93
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    • 2015
  • In 2013, the epidemics of pine wilt disease caused by the pine wood nematodes (Bursaphelenchus xylophilus) resulted in damages to the forests of black pine (Pinus thunbergii Parl.) trees in Jeju Island, Korea. Among the affected trees, an old black pine tree at Cave Temple on Mt. Sanbangsan was included and died due to the prevalence of pine wilt disease. The tree was on Mt. Sanbangsan, which was designated as a National Scenic Place with the Number 77 and was believed to be more than 400 years old in age. By examining the disc of the tree stem obtained from the height of 2 m, we counted the tree rings from 4 different directions and cross-dated the readings by comparing the records of drought simulated from the BROOK Model. Our analysis indicates that the tree seems to have grown since late 1860s. Contrary to the belief of the general public, we can conclude that the age of the tree was estimated to be at maximum 150 years, which means that it was not the same old tree as was shown in the painting of the Tam-Ra-Sun-Ryeok-Do (an old painting book for the Inspection Tour of Jeju Island) published in 1702. Discussion was extended to the life history of the tree in growth and leaning and the measures to protect the tree species from the damages of the pine wilt disease caused by pine wood nematodes.

Exploring Variables Related to Students' Understanding of the Convergence of Basic and Applied Science (기초과학과 응용과학의 융합에 관한 학생들의 이해와 관련된 변인 분석)

  • Ha, Min-Su;Lee, Jun-Ki
    • Journal of The Korean Association For Science Education
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    • v.32 no.2
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    • pp.320-330
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    • 2012
  • Understanding the relation between basic science and applied science is important for students in understanding the importance of learning science, the relationship between scientific knowledge and human life, and for enhancing their science motivation. In this study, we evaluated the statistical validity of this hypothetical model and explored the effect of gender and students' preferred courses (e.g., humanities, science, and art) on four dependent variables. We also evaluated the differences of students' understanding across scientific domains and students' understanding concerning basic and applied scientific knowledge. Three hundred and twenty five 10th grade students participated in this survey research. Statistically, we employed bivariate correlation, partial correlation, path analysis, two-way ANOVA, and repeated measures ANOVA. Our findings illustrated that our hypothetical model was statistically valid. In addition, the significant interaction effects of gender and students' preferred courses on each dependent variable were shown. Students have different levels of understanding of the convergence of basic and applied science, the relation between scientific knowledge and human life, and the importance of learning science across scientific domains (e.g., physics, chemistry, earth science and biology).

A Six-Layer SVAT Model for Energy and Mass Transfer and Its Application to a Spruce(Picea abies [L].Karst) Forest in Central Germany (독일가문비나무(Picea abies [L].Karst)림(林)에서의 Energy와 물질순환(物質循環)에 대(對)한 SLODSVAT(Six-Layer One-Dimensional Soil-Vegetation-Atmosphere-Transfer) 모델과 그 적용(適用))

  • Oltchev, A.;Constantin, J.;Gravenhorst, G.;Ibrom, A.;Joo, Yeong-Teuk;Kim, Young-Chai
    • Journal of Korean Society of Forest Science
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    • v.85 no.2
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    • pp.210-224
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    • 1996
  • The SLODSVAT consists of interrelated submodels that simulate : the transfer of radiation, water vapour, sensible heat, carbon dioxide and momentum in two canopy layers determined by environmental conditions and ecophysiological properties of the vegetation ; uptake and storage of water in the "root-stem-leaf" system of plants ; interception of rainfall by the canopy layers and infiltration and storage of rain water in the four soil layers. A comparison of the results of modeling experiments and field micro-climatic observations in a spruce forest(Picea abies [L].Karst) in the Soiling hills(Germany) shows, that the SLODSVAT can describe and simulate the short-term(diurnal) as well as the long-term(seasonal) variability of water vapour and sensible heat fluxes adequately to natural processes under different environmental conditions. It proves that it is possible to estimate and predict the transpiration and evapotranspiration rates for spruce forest ecosystems on the patch and landscape scales for one vegetation period, if certain meteorological, botanical and hydrological information for the structure of the atmospheric boundary layer, the canopy and the soil are available.

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Engraftment of Intraperitoneally Injected Bone Marrow Cells to Newborn Mice Injected with an Angiogenesis Inhibitor (혈관생성 억제제를 주사한 마우스 모델에서의 골수 세포의 복강 내 주입 후 생착)

  • Cho, Su-Jin;Ju, Sun-Young;Woo, So-Youn;Kang, Hyoung-Jin;Ahn, Hyo-Seop;Ryu, Kyung-Ha;Park, Eun-Ae
    • Neonatal Medicine
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    • v.15 no.1
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    • pp.22-31
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    • 2008
  • Purpose : Bronchopulmonary dysplasia (BPD) is characterized by arrested vascular and alveolar growth in the premature lung. Considering the consequences of arrested lung growth, the idea of administering bone marrow cells to enhance the inborn repair mechanism is promising as this may reduce the morbidity and mortality of BPD. We followed enhanced green fluorescent protein (EGFP)-labeled bone marrow cells (BMC) injected intraperitoneally into non-EGFP mice in order to determine their fate after transplantation. Methods : An angiogenesis inhibitor, SU1498, was injected subcutaneously on day 3 in non-EGFP C57BL/6 newborn mice to create a model of arrested alveolar development. On the following day, $1{\times}10^6$ BMCs isolated from major histocompatibility complex (MHC)- matched syngenic EGFP mice were injected intraperitoneally to non-EGFP BPD mice. Morphometric analysis, immunostaining, and confocal microscopy were performed to determine the fate of EGFP-positive stem cells in the injured lung. Results : SU1498 injection reduced alveolar surface area and mean alveolar volume in newborn mice. BMC injection resulted in recovery of lung structure comparable to controls. EGFP-positive BMCs were identified in the lungs of the recipient mice after intraperitoneal injection. The injected EGFP cells were co-stained with endothelial and epithelial cells of the developing lung as determined by confocal microscopy. Conclusion : Our results illustrated that EGFP-positive BMCs engrafted and trans-differentiated into epithelial and endothelial cells after intraperitoneal injection in a mouse model of arrested alveolar development.

Anti-inflammatory effect of Euonymi Lignum Suberalatum in LPS-activated Raw 264.7 cells and CA-induced paw edema rat model (귀전우(鬼箭羽)의 in vitro 및 in vivo에서의 염증억제효과)

  • Jeon, Chang Kwon;Park, Sang Mi;Park, Chung A;Byun, Sung Hui;Kim, Sang Chan
    • Herbal Formula Science
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    • v.27 no.2
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    • pp.101-120
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    • 2019
  • PURPOSE : Euonymi Lignum Suberalatum (EL) is the stem fin of Euonymi alatus. In traditional Korean medicine, EL is used for treatment of uterine bleeding, metritis and static blood. Recently, many studies have reported several pharmacological effects of EL including anticancer, antimicrobial, antidiabetic activity, and anti-oxidative stress. However, the mechanisms underlying anti-inflammatory effects by the EL is not established. METHODS : To investigate anti-inflammatory effects of Euonymi Lignum Suberalatum Water (ELWE), Raw 264.7 cells were pre-treated with $10-300{\mu}g/mL$ of ELWE, and then exposed to $1{\mu}g/mL$ of LPS. Levels of NO, IL-6, $IL-1{\beta}$ and $TNF-{\alpha}$ were detected by ELISA kit. Expression of pro-inflammatory proteins were determined by immunoblot analysis. To evaluate the anti-inflammatory effect in vivo, rat paw edema volume, and expressions of COX-2 and iNOS proteins in carrageenan (CA)-induced rat paw edema model. RESULTS : NO production activated by LPS, was decreased by $30-300{\mu}g/mL$ of ELWE. Production of inflammatory mediators such as $TNF-{\alpha}$, ILs, $PGE_2$ were decreased by ELWE 100 and $300{\mu}g/mL$. In addition, ELWE reduced LPS-mediated iNOS and COX-2 expression. Moreover, ELWE increased $I-{\kappa}B{\alpha}$ expression in cytoplasm and decreased $NF-{\kappa}B$ expression in nucleus. In vivo study, ELWE reduced the increases of paw swelling, and expression of iNOS and COX-2 proteins in paw edema induced by CA injection. CONCLUSION : The results indicate that ELWE could inhibit the acute inflammatory response, via modulation of $NF-{\kappa}B$ activation. Furthermore, inhibition of rat paw edema induced by CA is considered as clear evidence that ELWE may be a useful source to treat acute inflammation.

Attenuation of Experimental Autoimmune Hepatitis in Mice with Bone Mesenchymal Stem Cell-Derived Exosomes Carrying MicroRNA-223-3p

  • Lu, Feng-Bin;Chen, Da-Zhi;Chen, Lu;Hu, En-De;Wu, Jin-Lu;Li, Hui;Gong, Yue-Wen;Lin, Zhuo;Wang, Xiao-Dong;Li, Ji;Jin, Xiao-Ya;Xu, Lan-Man;Chen, Yong-Ping
    • Molecules and Cells
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    • v.42 no.12
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    • pp.906-918
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    • 2019
  • MicroRNA-223-3p (miR-223-3p) is one of the potential microRNAs that have been shown to alleviate inflammatory responses in pre-clinical investigations and is highly encased in exosomes derived from bone mesenchymal stem cells (MSC-exosomes). MSC-exosomes are able to function as carriers to deliver microRNAs into cells. Autoimmune hepatitis is one of the challenging liver diseases with no effective treatment other than steroid hormones. Here, we examined whether MSC-exosomes can transfer miR-223-3p to treat autoimmune hepatitis in an experimental model. We found that MSC-exosomes were successfully incorporated with miR-223-3p and delivered miR-223-3p into macrophages. Moreover, there was no toxic effect of exosomes on the macrophages. Furthermore, treatments of either exosomes or exosomes with miR-223-3p successfully attenuated inflammatory responses in the liver of autoimmune hepatitis and inflammatory cytokine release in both the liver and macrophages. The mechanism may be related to the regulation of miR-223-3p level and STAT3 expression in the liver and macrophages. These results suggest that MSC-exosomes can be used to deliver miR-223-3p for the treatment of autoimmune hepatitis.