• Title/Summary/Keyword: 동결배아

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TGF-$\alpha$로 분화 유도된 인간 배아줄기세포 이식에 따른 파킨슨 동물 모델 생쥐의 행동 개선

  • 이금실;김용식;신현아;조황윤;김은영;이원돈;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.271-271
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    • 2004
  • 본 실험은 TGF-a를 처리하여 분화가 유도된 인간배아 줄기세포를 파킨슨 동물모델에 이식하여 숙주세포에서의 생존 및 이식효과를 검토하고자 실시하였다. TGF-a로 분화된 세포의 이식효과를 판정하고자 배양시 TGF-a처리군과 처리하지 않은 군으로 나누어 분화를 유도한 인간배아 줄기세포를 hoechst33342로 표지 하여 병변 유발과 동일한 방법으로 동측 선조체내에 4×10⁴개/2ul가 되도록 이식하고(이식 위치: AP 0.7, ML 2.0, DV3.4) 이식 후 2, 4주에서 행동학적 변화를 관찰하고 4주에 동물을 희생시켜 4% PFA를 이용하여 뇌 조직을 고정하고 뇌 조직은 40㎛ 두께로 동결 절편을 만들어 면역조직화학염색을 시행하여 신경세포로의 분화 및 TH 발현 여부를 관찰하였고 분화의 표지물질로 nestin, NF200, GFAP, TH를 사용하여 형태학적 변화를 관찰하였다. (중략)

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Effects of Permeable Cryoprotectants on Viability of Mammalian Embryo Model (침투성 동결보호제가 포유류 초기배자의 생존성에 미치는 영향)

  • Kim, Hyun;Cho, Sang-Rae;Kim, Dong Kyo;Choe, Changyong;Seong, Hwan-Hoo
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.195-200
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    • 2015
  • The objective of this study was to evaluate the toxicities of permeable cryoprotectants and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Toxicities of permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), Glycerol, and 1,2-PROH were investigated using a murine embryo model. Female $F-{_1}$ mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to among DMSO, EG, Glycerol, and 1,2-PROH. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO and Glycerol at the 2-cell stage was significantly lower than that were treated with EG ($81.1{\pm}15.1$), 1,2-PROH ($88.0{\pm}21.1$) or the control ($99.9{\pm}21.3$) (p<0.001). On comparison of four cryoprotectant treated groups, the DMSO and Glycerol treated group showed a decreased cell count compared with the EG and 1,2-PROH treated group (p<0.05). Both DMSO ($14.7{\pm}1.3$), EG ($12.1{\pm}1.1$), Glycerol ($15.2{\pm}1.8$), and 1,2-PROH ($11.5{\pm}1.3$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.5{\pm}0.7$, p<0.0001). In addition, the DMSO or Glycerol treated group showed more apoptotic cells than the EG or 1,2-PROH treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to among DMSO, EG, Glycerol, and 1,2-PROH at room temperature. When comparing four permeable cryoprotective agents, EG and 1,2-PROH appeared to be less toxic than DMSO and Glycerol at least in a murine embryo model.

Consideration on Domestic Production of Materials and Consumables for Human IVF-ET Program (체외수정 및 배아이식술 관련 재료 및 소모품의 국산화 필요성에 대한 고찰)

  • Cha, Byung-Hun;Jun, Jin-Hyun
    • Development and Reproduction
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    • v.15 no.4
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    • pp.385-391
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    • 2011
  • Human in vitro fertilization and embryo transfer (IVF-ET) program is a general procedure for infertile couples since first successful delivery on 1978 in UK and Korean first on 1985. Recently in Korea, more than 42,000 cases per year of IVF-ET were performed and showed good pregnancy rates compared worldwide data. The human IVF-ET procedure use many consumables, such as ovum pick-up (OPU) needles, centrifuge tubes, culture dish, ICSI pipette, culture media and ET catheters. Major of these materials are supported by the global companies. Thus, Korean IVF-ET program might be placed unstable situation by global economical risks. These uncertain problems could be overcome by the domestic production of IVF-ET materials and consumables. Two times questionnaires for Korean clinicians and researchers about the domestic production were performed and analyzed. Many of them requested domestic OPU needles, ET catheters, culture media and ICSI pipettes under good quality control and quality assurance system. This trial may be contributed to industrialization and to global competence of Korean IVF-ET program. The results of this survey can be provide a fundamental base for development and production of domestic materials and consumables for human IVF-ET program.

Toxic Effects of Ethylene Glycol on Mammalian Embryo Survivability (Ethylene Glycol이 포유류 초기배자의 생존성에 미치는 독성 효과 분석)

  • Kim, Hyun;Yu, Dae Jung;Choe, Changyong;Seong, Hwan-Hoo
    • Reproductive and Developmental Biology
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    • v.39 no.3
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    • pp.77-81
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    • 2015
  • This study was carried out to evaluate the effects of embryonic stage and toxicities of cryoprotectant on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology. Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and ethylene glycol (EG) were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or EG. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO ($68.1{\pm}24.1$) at the 2-cell stage was significantly lower than that were treated with EG ($81.2{\pm}27.0$) or the control ($99.0{\pm}18.3$) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the EG treated group (p<0.05). Both DMSO ($15.4{\pm}1.5$) and EG ($10.2{\pm}1.4$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.1{\pm}0.9$, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the EG treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or EG at room temperature. When comparing two cryoprotective agents, EG appeared to be less toxic than DMSO at least in a murine embryo model.