• 제목/요약/키워드: Stem Cell

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원자간력현미경(AFM)을 이용한 줄기세포의 신경세포로 분화 인지에 관한 연구

  • 권상우;양우철;전송희;유보영;최윤경
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2012년도 제42회 동계 정기 학술대회 초록집
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    • pp.558-558
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    • 2012
  • 최근의 원자간력현미경(AFM)은 soft한 생체물질을 비파괴적 방법 및 나노크기의 분해능으로 여러 구조적, 물리적 특성 측정이 가능하여 bio분야에 다양이 활용되고 있다. 본 연구에서는 AFM을 이용하여 줄기세포인 BM MSC(bone marrow mesenchymal stem cell)가 신경세포로 분화 여부를 측정하는 방법을 보고하고자 한다. 신경세포의 신호전달은 시냅스에서 신경전달물질을 매개로 하여 이루어지는데, 신경전달물질 중에 D-Glutamic acid는 시냅스후세포에서 흥분성 전위 크기를 증가시킨 상태를 장기간 유지시켜주는 물질로, 특정물질인 Glutamate와 항원-항체 결합을 한다. 본 연구에서는 이 두 물질간의 항원-항체 반응을 활용하여 줄기세포의 신경세포로 분화 여부를 AFM으로 측정하였다. 먼저, 수용성 시료인 두 물질을 증류수에 용해시켜 Mica 기판에 그 용액을 떨어뜨려 자연건조로 시료를 준비한 후, AFM으로 형태 및 크기를 측정하였다. D-Glutamic acid와 Glutamate는 구형 입자 형태를 보였으며, Glutamate의 너비는 ~100 nm이고, D-Glutamic acid는 ~50 nm였다. 두 물질이 든 용액을 섞었을 때, 항원-항체 반응에 의해 다른 크기의 두 구형입자가 붙어 있는 형태가 관찰되었다. 이 반응을 활용하여, 신경세포에서 분비되는 신경전달물질인 D-Glutamic acid를 선별하였다. DMEM 배지에 신경암세포주인 SH-SY5Y 를 접종한 후 $37.6^{\circ}C$의 incubator에서 24시간 배양하고, 화학적 자극(60~70 mM의 KCl 용액을 주입함)을 주어 신경전달물질 분비를 유도하였다. 그 배지에 항체 Glutamate 를 주입하여 자연건조 시킨 후 항원-항체 결합특성을 AFM으로 측정하여, 항원-항체 결합된 이미지와 동일함을 확인하였다. 결과적으로 AFM을 이용한 신경전달물질의 항원-항체 결합여부 측정을 통해, BM MSC 줄기세포의 신경세포로 분화를 판단할 수 있으며, 이 방법은 줄기세포의 특정 세포로의 분화 여부 판단에 활용될 것으로 기대된다.

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Effect of Smoking and Alcohol Consumption on Seminal Quality in Young Mice

  • Park, Hyun-Jung;Choung, Eun-Hoi;Kim, Jae-Hwan;Chai, Seung-Youn;Choi, Jin-Yong;Kim, Jin-Hoi;Song, Hyuk
    • Reproductive and Developmental Biology
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    • 제33권4호
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    • pp.217-222
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    • 2009
  • The purpose of this study is to evaluate the effects of alcohol or cigarette smoking on seminal parameters in a large group of mice model. Nine groups (n=20/group) of mice were treated intensive noxious materials that abdominal injection of 21% (v/v) of ethanol, cigarette smoke (10, 20, 30 minutes/day), and combination of ethanol and 30 minutes of smoking. In addition, vitamin C and selenium were also treated to mice exposed to combination of alcohol and smoking to identify the recovering effect. Sperm viability and motility were significantly decreased in either alcohol consumption or smoking exposed group, and combination of both materials have additive detrimental effects on seminal parameters. Mice groups that exposed to alcohol and smoking showed statistically significant decrease in motility and increase of static spermatozoa. Moreover, combination of both treatments showed cumulative effect in increase of static spermatozoa. Treatments of either vitamin C or selenium dramatically recovered detrimental effects of alcohol and smoking on seminal quality, although combination of both antioxidant molecules did not show any additive effect. In conclusion, detrimental effects of alcohol and cigarette consumption on sperm quality and motility were identified in mice model, and these detrimental effects can be compensated to uptake of anti-oxidant molecules.

Gd-doped $CeO_2$ 와 Sr-doped $LaMnO_3$ 분말의 합성 및 그 계면에서의 상 안정성 연구 (A Study on the Synthesis of Gd-doped $CeO_2$ and Sr-doped $LaMnO_3$ Powders and Phase Stability in Their Interface)

  • 정승훈;김남진;이덕열
    • 한국세라믹학회지
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    • 제34권6호
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    • pp.652-658
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    • 1997
  • The phase stability in the interface of Sr-doped LaMnO3(LSM)/Gd-doped CeO2(CGO) was examined in this study in order to check the feasibility of using LSM as the cathode material in a low-temperature SOFC(solid oxide fuel cell) using CGO as the electrolyte. For the purpose, CGO powders of Ce0.82Gd0.18O0.91 and two LSM powders having different compositions, La0.9Sr0.1MnO3(LSM10) and La0.5Sr0.5MnO3(LSM50), were synthesized using Pechini method. Then, specimens having the LSM/CGO interface were prepared, heat-treated at 130$0^{\circ}C$ for up to 3 days, and analyzed by XRD and STEM/EDX. Face-centered cubic CGO powders of less than 10 nm size were obtained by calcination of polymeric precursor formed in the process at 45$0^{\circ}C$. Higher calcination temperature of $700^{\circ}C$ was necessary for monoclinic LSM10 and cubic LSM50 powders. LSM powders were coarser than CGO and observed to be in the range of 50~100 nm. No trace of LSM-CGO interaction product was found in the XRD pattern. Also it was known from the concentration profile in the vicinity of the interface that interdiffusion was occurred over only a small penetration depth of ~100 nm order.

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Sperm chromatin and DNA integrity, methyltransferase mRNA levels, and global DNA methylation in oligoasthenoteratozoospermia

  • Rahiminia, Tahereh;Yazd, Ehsan Farashahi;Fesahat, Farzaneh;Moein, Mohammad Reza;Mirjalili, Ali Mohammad;Talebi, Ali Reza
    • Clinical and Experimental Reproductive Medicine
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    • 제45권1호
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    • pp.17-24
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    • 2018
  • Objective: To investigate sperm chromatin/DNA integrity, global DNA methylation, and DNMT mRNA transcription in men with oligoasthenoteratozoospermia (OAT) compared with normozoospermic men. Methods: Semen samples from 32 OAT patients who comprised the case group and 32 normozoospermic men who comprised the control group were isolated and purified using a standard gradient isolation procedure according to World Health Organization criteria. DNMT1, DNMT3A, and DNMT3B transcripts were then compared between groups using real-time quantitative reverse-transcription polymerase chain reaction. Global DNA methylation in sperm was determined by an enzyme-linked immunosorbent assay. Protamine deficiency and the proportion of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3), aniline blue (AB), and toluidine blue (TB) staining, as well as the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The p-values < 0.05 were considered to indicate statistical significance. Results: Significantly higher proportions of AB+, TB+, CMA3+, and TUNEL+ spermatozoa, as well as DNMT3A and DNMT3B transcription, were found in the OAT group. Positive correlations were detected between sperm parameters, DNA/chromatin damage, and DNMT3A and DNMT3B transcripts. Global DNA methylation was significantly higher in the OAT patients and had a significant correlation with abnormal results of all sperm chromatin integrity tests, but was not associated with DNMT1, DNMT3A, or DNMT3B expression. Conclusion: Oligoasthenoteratozoospermic men showed abnormal sperm parameters, abnormal chromatin/DNA integrity, and a higher global DNA methylation rate, as well as overexpression of DNMT mRNA.

Ricinus communis extract inhibits the adipocyte differentiation through activating the Wnt/β-catenin signaling pathway

  • Kim, Bora;Kim, Hyun-Soo
    • 한국식품저장유통학회지
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    • 제24권4호
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    • pp.524-528
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    • 2017
  • Ricinus communis, belongs to the family Euphorbiaceae, has been known as medicinal plants for treatment of inflammation, tumors, antidiabetic, hepatoprotective and laxative. Compared to many pharmacological studies, the effect of R. communis extract on regulating adipogenesis as therapeutic drug for treating obesity has not been reported. R. communis extract (RCE) was investigated to determine its effects on the adipogenesis by monitoring the status of $Wnt/{\beta}-catenin$ signaling and factors involving the differentiation of adipocytes. The differentiation of 3T3-L1 cells monitored by Oil Red O staining was inhibited in concentration dependent manner by RCE. The luciferase activity of HEK 293-TOP cells containing pTOPFlash with Tcf4 response element-luciferase gene was increased approximately 2-folds by the treatment of RCE at concentrations of $100{\mu}g/mL$ compared to the control. Activation of the $Wnt/{\beta}-catenin$ pathway by RCE was further confirmed by immunocytochemical analysis which shows an increment of nuclear localization of ${\beta}-catenin$. In addition, safety of RCE was verified through performing neural stem cell morphology assay. Among the identified flavonoids in RCE, isoquercitrin was the most abundant. Therefore, these results indicate that the adipocyte differentiation was significantly reduced by isoquercitrin in R. communis. In this study, RCE suppresses the adipogenesis of 3T3-L1 cells via the activation of $Wnt/{\beta}-catenin$ signaling.

혈액종양 관련 국내 간호연구 분석 (Analysis of Research on the Nursing of Hematology in Korea)

  • 김형순;반자영;윤지연;나영희;전진영;여순미;유지연
    • 종양간호연구
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    • 제10권2호
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    • pp.146-155
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    • 2010
  • Purpose: This study was conducted in order to analyze themes, concepts, research methods and results of previous domestic research on the nursing of hematologic patients conducted through the last 10 yr, to find trends in the research, and to provide basic materials for setting the direction of future research on the nursing of hematologic patients. Methods: This study analyzed a total of 72 nursing theses related to hematology sampled from domestic theses for a master's or doctoral degree and papers published in six nursing journals registered in Korea Research Foundation from January 2000 to July 2009. Results: Of the 72 theses, 51 were for a master's degree, 7 for a doctoral degree, and 14 not for an academic degree. The concept covered most frequently in correlation research was 'quality of life' and concepts found in comparative research were stress and quality of life. In experimental research, the most common nursing intervention was oral care. The scale used most frequently was Spielberger's Anxiety Scale. Conclusion: It is necessary to expand experimental research applying nursing interventions, and to use objective physiological indexes for more effective assessment in experimental research. Furthermore, interdisciplinary research is required for enhancing the quality of clinical nursing research.

The treatment of pediatric chronic myelogenous leukemia in the imatinib era

  • Lee, Jae-Wook;Chung, Nack-Gyun
    • Clinical and Experimental Pediatrics
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    • 제54권3호
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    • pp.111-116
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    • 2011
  • Childhood chronic myelogenous leukemia (CML) is a rare hematologic disease, with limited literature on the methods of treatment. Previously, allogeneic hematopoietic stem cell transplantation (HSCT) was considered the only curative treatment for this disease. Treatment with imatinib, a selective inhibitor of the BCR-ABL tyrosine kinase (TKI), has resulted in prolonged molecular response with limited drug toxicity. Imatinib is now implemented in the primary treatment regimen for children, but the paucity of evidence on its ability to result in permanent cure and the potential complications that may arise from long-term treatment with TKIs have prevented imatinib from superseding HSCT as the primary means of curative treatment in children. The results of allogeneic HSCT in children with CML are similar to those observed in adults; HSCT-related complications such as transplant-related mortality and graft-versus-host disease remain significant challenges. An overall consensus has been formed with regards to the need for HSCT in patients with imatinib resistance or those with advanced-phase disease. However, issues such as when to undertake HSCT in chronic-phase CML patients or how best to treat patients who have relapsed after HSCT are still controversial. The imatinib era calls for a reevaluation of the role of HSCT in the treatment of CML. Specific guidelines for the treatment of pediatric CML have not yet been formulated, underscoring the importance of prospective studies on issues such as duration of imatinib treatment, optimal timing of HSCT and the type of conditioning utilized, possible treatment pre-and post-HSCT, and the role of second-generation TKIs.

Clinical manifestations of BK virus infection in pediatric kidney transplant patients

  • Kwon, Yiyoung;Kim, Jeong Yeon;Lee, Yeonhee;Cho, Heeyeon
    • Clinical and Experimental Pediatrics
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    • 제62권11호
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    • pp.422-427
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    • 2019
  • Background: Polyomavirus BK (BKV) infection is an important cause of graft loss in kidney transplant patients. Purpose: The purpose of this study was to evaluate clinical findings and risk factors for BKV in pediatric patients after kidney transplantation. Methods: This retrospective single-center study included 31 pediatric kidney transplant recipients from January 2002 to December 2017. Two patients received 2 transplantations during the study period, and each transplant was analyzed independently. Total number of cases is 33 cases with 31 patients. BKV infection was confirmed from blood samples via periodic quantitative polymerase chain reaction. Results: The mean age at kidney transplantation was 11.0±4.7 years, and the male-to-female ratio was 2.7:1. Three patients had a past medical history of high-dose chemotherapy and autologous stem-cell transplantation for solid tumors. Nine patients (27.3%) developed BKV infection. The median period from kidney transplantation to BKV detection in blood was 5.6 months. There was no statistically significant difference in estimated glomerular filtration rate between patients with and those without BKV infection. Among 9 patients with BKV viremia, 7 were treated by reducing their immunosuppressant dose, and BKV was cleared in 6 of these 7 patients. In the other 2 BKV-positive patients, viremia improved without immunosuppressant reduction. Conclusion: BKV infection is common in children with kidney transplantation and might not have affected short-term renal function in our patient sample due to early immunosuppressant reduction at the time of BKV detection.

급성 골수성 백혈병 환자에서 G-CSF를 포함한 고용량 화학요법 후 자가 말초혈 조혈모세포이식의 효과 (Autologous Peripheral Blood Stem Cell Transplantation Using G-CSF Combined Conditioning in AML Patients)

  • 김병수;국훈;황태주;최철원;김준석
    • IMMUNE NETWORK
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    • 제2권1호
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    • pp.49-52
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    • 2002
  • Background: The possibility that G-CSF recruits leukemic cells from the G0 to S phase, which may lead to a greater susceptibility to cytotoxic drugs, such as ara-C, has been presented in Harada's study. Methods: In this study, we referred to the protocol of Harada et al 1 to try G-CSF combined marrow-ablative chemotherapy and autologous PBSCT, for the treatment of AML patients in CR1 status. Between January 1997 and March 1998, six AML patients (3: children, 3: adults) in CR1 status were autografted and followed up to 3 years. Results: The major regimen related toxicity was composed of mucositis and diarrhea without death. The time of ANC recovery to 500/L and 1,000/L was 11~48 and 16~81 days, respectively. The mean time of platelet recovery to 20,000/L and 50,000/L was 21~233 and 35~370 days, respectively. The platelet recovery time to 50,000/L was markedly prolonged for more than 100 days in four patients (66.7%). Moreover, four patients (66.7%) experienced a relapse of leukemia after transplantation, with a mean interval of 147.5 days after PBSCT. Two patients were in CR status for 53 and 51 months after PBSCT, respectively. Conclusion: The G-CSF combined marrow-ablative chemotherapy and autologous PBSCT resulted in a markedly delayed platelet recovery and no advantages for decreasing the relapse rate of AML. But, further studies will be warranted.

Effects of Pre-conditioning dose on the Immune Kinetics and Cytokine Production in the Leukocytes Infiltrating GVHD Tissues after MHC-matched Transplantation

  • Choi, Jung-Hwa;Yoon, Hye-Won;Min, Chang-Ki;Choi, Eun-Young
    • IMMUNE NETWORK
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    • 제11권1호
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    • pp.68-78
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    • 2011
  • Background: Graft-versus-host disease (GVHD) is a huddle for success of hematopoietic stem cell transplantation. In this study, effects of irradiation dose on immune kinetics of GVHD were investigated using B6 ${\rightarrow}$ BALB.B system, a mouse model for GVHD after MHC-matched allogeneic transplantation. Methods: BALB.B mice were transplanted with bone marrow and spleen cells from C57BL/6 mice after irradiation with different doses. Leukocytes residing in the peripheral blood and target organs were collected periodically from the GVHD hosts for analysis of chimerism formation and immune kinetics along the GVHD development via flow cytometry. Myeloid cells were tested for production of IL-17 via flow cytometry. Results: Pre-conditioning of BALB.B hosts with 900 cGy and 400 cGy resulted in different chimerism of leukocytes from the blood and affected survival of GVHD hosts. Profiles of leukocytes infiltrating GVHD target organs, rather than profiles of peripheral blood leukocytes (PBLs), were significantly influenced by irradiation dose. Proportions of IL-17 producing cells in the infiltrating $Gr-1^+$ or $Mac-1^+$ cells were higher in the GVHD hosts with high does irradiation than those with low dose irradiation. Conclusion: Pre-conditioning dose affected tissue infiltration of leukocytes and cytokine production by myeloid cells in the target organs.