• 제목/요약/키워드: Human amniotic fluid

검색결과 32건 처리시간 0.028초

인간 $\alpha$-fetoprotein에 대한 모노클로날 항체의 제조 및 모노클로날 항체를 이용한 효소면역분석법의 개발 (Production of a Monoclonal Antibody to Human $\alpha$-Fetopotein and Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assays for Human $\alpha$-Fetoprotein)

  • Michung Yoon;Hyun-Hee Lee;Youngwon Lee
    • 대한의생명과학회지
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    • 제5권1호
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    • pp.1-10
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    • 1999
  • 본 연구에서는 혈장이나 양수에 있는 $\alpha$-fetoprotein (AFP)을 인식 할 수 있는 모노클로날 항체를 제조하고, 모노클로날 항체를 이용한 효소면역분석법을 개발하고자 하였다. 양수로부터 얻은 AFP를 쥐에 주사한 후 비장을 분리하여 종양세포 (Sp2/O-Ag-14)와 융합하였고, 하이브리도마 기술을 이용하여 모노클로날 항체를 제조하였다. 모노클로날 항체를 클로닝하였으며, 생성된 항체를 MabF22로 명명하였고, IgG1 중사슬과 k 경사슬의 isotype을 나타냈다. 또한 immunoblotting 방법과 ELISA로 특이도를 조사한 결과 모노클로날 항체는 AFP와만 반응하였고, 결합 친화상수는 0.8$\times$$10^{-10}$M이었다. 두 종류의 효소면역분석법 -경쟁적 또는 비경쟁적 분석 -을 이용하여 항체의 효용성을 조사하였으며, 두 방법 모두 AFP와 농도에 비례하여 반응하였다. 따라서 본 연구에서 생산된 모노클로날 항체는 연구목적으로 뿐만 아니라 AFP 농도를 측정하기 위한 면역진단시약의 개발에도 유용할 것으로 생각된다.

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인간 양수 유래 줄기세포에서 세포투과단백질을 이용한 ESRRB 단백질의 직접도입에 의한 전분화능 관련 유전자의 발현 조절 (Up-regulation of Pluripotency-related Genes in Human Amniotic Fluid-derived Stem Cells by ESRRB Conjugated with Cell-Penetrating Peptide)

  • 조중현;이유선;오미희;고정재;전용필;이동률
    • 한국발생생물학회지:발생과생식
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    • 제14권4호
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    • pp.243-251
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    • 2010
  • Estrogen related receptor $\beta$(Esrrb)는 오르판 수용체 중 하나로 전분화능 관련유전자인 Oct4와 Nanog의 발현을 조절함으로써 줄기세포의 미분화를 유지시키고, 지속적인 자기 복제를 가능케 하는 유전자로 알려져 있다. 또한 Feng 등 (2009)은 체세포에 Oct4, Sox2와 함께 Esrrb 유전자를 함께 도입하면, 유전자가 변형된 체세포가 배아 줄기세포와 유사한 유도만능줄기세포로 리프로그래밍(reprograming)되어 진다는 결과를 보고한 바 있다. 본 연구에서는 인간 ESRRB 단백질을 양수유래줄기세포 내로 직접도입하는 방법을 개발하고, 이를 통해 전분화능 관련유전자의 기능 조절을 확인하고자 하였다. 클로닝 된 인간 short-form ESRRB를 세포투과 펩타이드(cell-penetrating peptide, CPP)의 일종인 R7(아르기닌 7개)에 접합(Fusion)하였고, 합성단백질 (R7-ESRRB-His6)의 형태로 배양중인 인간 양수 유래 줄기세포에 처리하여 세포내로 도입하였다. R7-ESRRB-His6 단백질은 5시간 내에 세포막을 통과하였고, 24시간 내에 핵 내로 이동하였다. 또한 핵 내로 이동한 ESRRB 단백질은 OCT4와 NANOG 유전자의 발현을 증가시켰을 뿐만 아니라, 또 다른 전분화능 관련유전자인 SOX2의 발현도 함께 증가시킨다는 것을 확인하였다. 이상의 결과는 세포투과 펩타이드와 유전자의 접합을 통해 생산된 R7-ESRRB-His6 합성단백질이 양수유래줄기세포내로 원활하게 도입되는 것을 확인하였고, 유전자의 변형 없이 전분화능 관련유전자의 기능을 조절할 수 있는 방법임을 확인하였다.

Constructing intelligent agent for chromosome knowledge base

  • Shin, Yong-Won
    • 한국산학기술학회:학술대회논문집
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    • 한국산학기술학회 2003년도 Proceeding
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    • pp.3-9
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    • 2003
  • The task for chromosome analysis and diagnosis by experienced cytogenetists are being concerned as repetitive, time consuming job and expensive. For that reason, intelligent agent based on chromosome knowledge base has been established to be able to analyze chromosomes and obtain necessary advises from the knowledge base instead of human experts. That is to say, knowledge base by IF THEN production rule was implemented to a knowledge domain with normal and abnormal chromosomes, and then the inference results by knowledge base could enter the inference data into the database. Experimental data were composed of normal chromosomes of 2,736 patients 'cases and abnormal chromosomes of 259 patients' cases that have been obtained from GTG-banding metaphase peripheral blood and amniotic fluid samples. The completed intelligent agent for chromosome knowledge base provides variously morphological information by analysis of normal or abnormal chromosomes and it also has the advantage of being able to consult with user on chromosome analysis and diagnosis.

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지식 베이스를 이용한 교육용 염색체 분석 시스템 (Chromosome Analysis System based on Knowledge Base for CAI)

  • 박정선;신용원
    • 한국지능정보시스템학회:학술대회논문집
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    • 한국지능정보시스템학회 2001년도 춘계정기학술대회
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    • pp.215-222
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    • 2001
  • The task for chromosome analysis and diagnosis by experienced cytogenetists are being concerned as repetitive, time consuming job and expensive. FOr that reason, chromosome analysis system based on knowledge base for CAI had been established to be able to analyze chromosomes and obtain necessary advises from the knowledge base instead of human experts. That s to say, knowledge base by IF THEN production rule was implemented to a knowledge domain with normal and abnormal chromosomes, and then the inference results by knowledge base could enter the inference data into the database. Experimental data were composed of normal chromosome of 2,736 patients'cases and abnormal chromosomes of 259 patients'cases that have been obtained from GTG-banding metaphase peripheral blood and amniotic fluid samples. The complete system provides variously morphological information by analysis of normal or abnormal chromosomes and it also has the advantage of being able to consult with user on chromosome analysis and diagnosis.

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성숙배양액에 첨가하는 인간체액 (Human Body Fluids) 및 성선자극호르몬이 생쥐 미성숙난자의 핵성숙과 수정능력에 미치는 영향 (Influences of Human Body Fluids and Gonadotropins Supplemented in the Maturation Medium on the Nuclear Maturation and Fertilizability of Mouse Immature Oocytes)

  • 박기상;손원영;김진희;이경아;한세열;고정재;차광열
    • Clinical and Experimental Reproductive Medicine
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    • 제21권2호
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    • pp.183-190
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    • 1994
  • Purpose of the present study was to find the optimal culture conditions for the maturation and fertilization of immature oocytes by the use human body fluids and gonadotropins (Gn) in the mouse model. Cumulus-enclosed mouse immature oocytes were incubated in the medium containing various human body fluids with or without Gn in vitro, and examined to confirm nuclear maturation (NM) and fertilization. Female ICR mice were stimulated with 7.5 IU pregnant mares' serum gonadotropin (PMSG). Cumulus-enclosed immature oocytes were isolated at 48-52 hr post PMSG injection and cultured in TCM 199 supplemented with various concentrations (20, 50, and 70%) of human body fluids such as fetal cord serum (hCS), follicular fluid (hFF), peritoneal fluid (hPF) and amniotic fluid (hAF) in the presence or absence of 10 IU/ml PMSG and 10 IU/ml human chorionic gonadotropin (hCG) for 18 hr. Fetal calf serum (FCS) was used as a control for the supplements. Matured oocytes were fertilized with sperm collected from the epididymis of male mice. Fertilization was conducted in T6 medium containing 15 mgl ml bovine serum albumin, and confirmed at 6 hr post-insemination. Evaluation of nucler maturation and fertilization was carried out by rapid staining using fuchin. There was no significant difference between the effects of human body fluids and FCS supplements on nuclear maturation of cumulus enclosed mouse immature oocytes. When maturation medium was supplemented with 20% hPF or 20% hAF, fertilization rates were significantly (P<0.01) lower than that of 20% FCS, hCS and hFF groups. However, higher concentrations of body fluids during IVM were not more beneficial on fertilizability of oocytes. The addition of Gn significantly increased the fertilization rates in hPF and hAF groups (hPF without Gn; 51.5%, compared with 85.1% for addition of Gn, and hAF without Gn; 30.1% compared with 85.8% for addition of Gn) at 20% concentration. These results suggest that human body fluids at 20% concentration and gonadotropins can be used as supplements for the maturation of mouse immature oocytes in vitro. When gonadotropins supplemented with the human body fluids in the maturation medium, fertilizability of mouse immature oocytes was increased in hPF and hAF groups. These results can be applied to maturation of human immature oocytes in vitro.

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Whitten 배양액내 인간양수의 첨가가 생쥐 수정란의 체외발달 및 체외신장에 미치는 영향 (Effects of Human Amniotic Fluid Supplemented to Whitten's Medium on Development and Outgrowth of Mouse Embryo)

  • 김재환;승경록;최종현;정두용;이훈택;정길생
    • 한국가축번식학회지
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    • 제18권2호
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    • pp.87-94
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    • 1994
  • 본 연구는 생쥐 초기배 및 부화후 체외신장에 미치는 인간 양수의 첨가효과를 조사하고자 실시하였다. 생쥐 수정란의 체외발달에 미치는 인간 양수의 적정농도를 확인하고자 기본배양액 (Whitten's medium)에 16주령의 양수 농도를 각각 달리하여 체외배양을 실시한 결과, 20%를 첨가한 구에서 가장 높은 배반포로의 발달율과 부화율을 나타내었으며, 그 이상의 농도에서는 배발달율이 저하되었다. 포유동물 수정란 체외배양시 가장 많이 사용되는 첨가제인 fetal calf serum (FCS)과 bovine serum albumin(BSA)을 첨가한 구화의 배발달 성적을 비교한 결과, 임신중기 양수를 20% 첨가한 구의 배반포로의 발달율 (92.8%)과 부화율 (75.7%)은 기본배양액에서 배양된 것보다 배반포로의 발달율 (82.8%)과 부화율 (31.3%)은 높았으나 0.3% BSA (90.5%, 70.8%)나 10% FCS 첨가구 (94.3%, 74.3%)와는 유의한 차이가 인정되지 않았다. 임신주령에 따른 양수의 첨가효과를 조사한 결과 20%의 임신말기 양수를 첨가한 구의 배반포로의 발달율 (71.9%)과 부화율 (57.3%)은 20% 임신중기 양수를 첨가한 구보다 낮았으며, 부화후 배의 체외신장은 임신중기 양수와 FCS이 첨가된 구에서는 유기되었으나, 임신말기 양수와 BSA가 첨가된 구에서는 배의 체외신장이 유기되지 않았다. 이상의 본 연구결과를 통해 임신중기 양수 내에는 배발달 촉진인자와 배의 체외신장을 유기시키는 물질이 함유되어 있어 포유동물 배의 체외배양에 상당한 영향을 미치고 있다고 사료된다.

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한국 한의서를 중심으로 살펴본 '비훈(鼻熏)요법'의 활용에 대한 연구 (A Study on the Use of Bihoon (鼻熏) Therapy, which focuses on Korean traditional medicine)

  • 김동율;이지영
    • 한국의사학회지
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    • 제36권2호
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    • pp.99-113
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    • 2023
  • This paper is a study to find the philological basis of Bihoon (鼻熏) therapy. There is no separate philological study of Bihoon therapy to date, and for this reason, there is no clear definition or specific treatment manual. In this study, a related database was created and analyzed by examining literature data related to Bihoon therapy, focusing on Korean traditional medical books. There were about 1,000 data points related to Bihoon therapy in 45 kinds of medical books. They were largely classified into 1. Acute diseases such as insensitivity, 2. Diseases that occur in the upper human body such as nose, head, eyes, and throat, 3. Women's diseases related to childbirth, 4. Treatment of skin diseases and prevention of infectious diseases. In the case of insensitivity treatment, the focus was on awakening the patient's mind, and the treatment of diseases such as the nose, head, eyes, etc. was focused on resolving each symptom. Symptoms related to childbirth were mainly treated for uterine escapism or fainting after childbirth, while skin diseases were mainly treated for diseases that did not heal well, such as amniotic fluid. If a multifaceted approach to non-discipline therapy is added in the future, it is expected that clinical utilization will also be increased.

중간엽줄기세포의 초자화 동결법에 의한 냉동보존 (Cryopreservation of Mesenchymal Stem Cells by Vitrification)

  • 이효종;강선영;박세진;이승용;이희천;고필옥;박지권;백원영;연성찬
    • 한국임상수의학회지
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    • 제28권4호
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    • pp.394-398
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    • 2011
  • Mesenchymal stem cells (MSC) are pluripotent cells that can be found in umbilical cord blood from new borne babies as well as placenta, bone marrow, adipose tissue, amniotic fluid, muscle, et al. MSC are capable of renewing themselves without differentiation in long-term culture, also can be differentiated into various tissues under specific condition. Formulating a cryopreservation protocol for the MSC is required because these cells cannot survive for long periods under in vitro culture conditions and a new formulation of harmless cryoprotectant is needed for the direct injection of MSC into patients. The undifferentiated MSC were frozen with a vitrification solution of 40% ethylene glycol, 20% Ficoll-70 and 0.3M sucrose. The survival rate after thawing and their proliferation rate were examined and compared with slow rate cooling methods using dimethylsulfoxide (DMSO). The vitrification method showed high survival rate after thawing and proliferation capacity comparable to DMSO. It can be suggested that ultra-rapid cooling method by vitrification is reliable methods for long term preservation of MSC and the vitrification solution with ethylene glycol, Ficoll-70 and sucrose will be more beneficially used for direct transplantation of MSC into patients than DMSO solution.