• Title/Summary/Keyword: frozen cell

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Effect of Ethylene Glycol(EG) and Propylene Glycol(PG) on the Viability of Frozen-thawed Primordial Germ Cells(PGCs) on Korean Native Chicken(Ogye) by Vitrification (한국재래닭(오계)의 유리화 동결 시 생존율에 미치는 Ethylene Glycol(EG)과 Propylene Glycol(PG)의 영향)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Choi, Sung Bok;Ko, Yeoung Gyu;Do, Yoon Jung;Seong, Hwan-Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.197-205
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    • 2013
  • This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

Analysis of Semen Parameters, Sperm Activity, and Fertility of Somatic Cell Cloned Hanwoo Bulls (체세포 복제 한우 수소의 정액 성상, 정자의 활동성 및 수정 능력 분석)

  • Bae, Seong-Hoon;Hwang, Seong-Soo;Yang, Byong-Chul;Go, Yeoung-Kyu;Kim, Dong-Hoon;Im, Gi-Sun;Choi, Hwa-Sik;Jin, Dong-Il;Yang, Boh-Suk;Seong, Hwan-Hoo
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.139-143
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    • 2007
  • This study was performed to investigate the reproductive characteristics of the cloned Hanwoo bulls produced by SCNT. The semen ejaculated from the cloned bulls (C-38 and C-39) and normal Hanwoo bull was properly measured the volume, the number of sperm, and the viability of frozen-thawed sperm. The sperm activity was analyzed using computer assisted sperm analysis (CASA). To analyze fertilizing ability of the cloned bulls, in vitro fertilization and artificial insemination were performed using the frozen-thawed semen. There were no differences in semen volume, sperm concentration, and the viability of frozen-thawed sperm between cloned bulls and normal bull. The difference was statistically significant in total motility, curvilinear velocity (VCL), straight-line velocity (VSL), and average-path velocity (VAP) of both cloned bulls compared to those of normal Hanwoo bull, respectively (p<0.05). The cleavage and blastocyst development rate were not different between the groups. five cloned cows were artificially inseminated using the frozen-thawed semen of C-38, two of them became pregnant. Two second generation calves (one male and one female) were produced. Based on these results, the cloned Hanwoo bulls showed normal reproductive abilities of semen parameters and sperm activity to their comparators and produced cloned calves, although there are some individual differences on the parameters.

Live birth in a woman with recurrent implantation failure and adenomyosis following transfer of refrozen-warmed embryos

  • Safari, Somayyeh;Faramarzi, Azita;Agha-Rahimi, Azam;Khalili, Mohammad Ali
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.3
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    • pp.181-184
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    • 2016
  • The aim was to report a healthy live birth using re-vitrified-warmed cleavage-stage embryos derived from supernumerary warmed embryos after frozen embryo transfer (ET) in a patient with recurrent implantation failure (RIF). The case was a 39-year-old female with a history of polycystic ovarian syndrome and adenomyosis, along with RIF. After ovarian hyperstimulation, 33 cumulus-oocyte complexes were retrieved and fertilized with conventional in vitro fertilization and intracytoplasmic sperm injection. Because of the risk of ovarian hyperstimulation syndrome, 16 grade B and C embryos were vitrified. After 3 and 6 months, 3 and 4 B-C warmed embryos were transferred to the uterus, respectively. However, implantation did not take place. Ten months later, four embryos were warmed, two grade B 8-cell embryos were transferred, and two embryos were re-vitrified. One year later, the two re-vitrified cleavage-stage embryos were warmed, which resulted in a successful live birth. This finding showed that following first warming, it is feasible to refreeze supernumerary warmed embryos for subsequent ET in patients with a history of RIF.

PROPERTY CHANGES OF POLYMER ELECTROLYTE MEMBRANES WITH FREEZE/THAW CYCLES (동결/해동 조건에서 고분자막의 특성 변화 연구)

  • Park Gu-Gon;Lim Nam-Yun;Sohn Young-Jun;Park Jin-Soo;Lee Won-Yong;Kim Sae-Hoon;Lim Tae-Won;Kim Chang-Soo
    • 한국신재생에너지학회:학술대회논문집
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    • 2005.06a
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    • pp.281-283
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    • 2005
  • Water management in polymer electrolyte membrane fuel cells(PEMFCs) is one of the most challenging issues. Freeze start-up in the automotive applications is also important research topic in the PEMFC field. Transportation of proton and separation of reactant gases are main roles of polymer electrolyte membranes. It has been known that water in the membrane conducts as a vehicle for the proton transportation. At sub-zero temperature, the frozen water blocks the access of reactant gases to the active sites of electrode as well as occurs the physical destruction of fuel cell structures. In this study, property changes of electrolyte membranes in the freeze conditions $(at\;-25^{\circ}C)$ were investigated. For the various amount of water contained membranes, the property changes, especially for the proton conductivity, were observed after several times of freeze/thaw$(-25\~80^{\circ}C)$ cycle.

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Production of monoclonal antibody(MAb) for lymphocytes of Korean native cattle 1. MAb to T lymphocytes (한우 림프구에 대한 단일항체 생산 1. T 림프구에 대한 단일항체)

  • Song, Hee-jong;Woon, Jae-ho;Kim, Tae-joong;Chai, Hyo-seok;Hur, Boo-hong;Choi, Min-soon
    • Korean Journal of Veterinary Research
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    • v.35 no.4
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    • pp.769-776
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    • 1995
  • This study was undertaken to develop the monoclonal antibody(MAb) for lymphocytes of Korean native cattle by the cell hybridization of myeloma P3/NS-1/ 1-Ag-4-1 and spleen cells from BALB/c mice hyperimmunized with nylon wool column eluted peripheral T lymphocytes of Korean native cattle. The isotype of MAb KCT-14 against T lymphocyte was mouse $IgG_1$. KCT-14 positivity of mononuclear cells(MNC) from peripheral blood lymphocytes, nylon wool nonadherent and adherent-lymphocytes was 41.7%, 58.4% and 22.6%, respectively. And that of mesenteric lymph node-, spleen and thymus-MNC was 43.3%, 40.2% and 33.6%, respectively. Immunoperoxidase staining of frozen tissue sections showed that the MAb positive cells were located in the medulla of the thymus and in the paracortical area and the mantle zone of the germinal center in the lymph nodes. These results indicated that KCT-14 was one of the MAb for investigate of T lymphocyte subpopulations in the Korean native cattle.

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A Case Report of Lymphangioma Circumscriptum on Scalp (두피에 발생한 국한성 림프관종의 치험례)

  • Lee, Dong-Gwan;Shin, Hea-Kyeong;Seoul, Jung-Hyun;Choi, Jun
    • Archives of Craniofacial Surgery
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    • v.9 no.1
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    • pp.38-40
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    • 2008
  • Purpose: Lymphangioma circumscriptum is a rare, congenital benign hamartous malformation, caused by the saccular dilatation of lymph channels lines by normal, single cell, lymphatic endothelia that present as local eruptions of persistent, grouped, translucent vesicles. The lymphangioma circumscriptum lesions may occur on axillary fold shoulder, neck proximal limbs and buccal mucosa. We reported a rare case of lymphangioma circumscriptum on the scalp. Methods: A 15-year-old girl with a $5{\times}3cm$ sized lymphangioma circumscriptum on scalp was examined. It was defined a boundry by ultrasound. And then, a tissue crescent type expander with 120 cc normal saline was inserted on occipital area for a month. After confirmed safety margin of the excised lymphangioma circumscriptum on frozen biopsy and the scalp flap was elevated and covered with empty space. Results: A histopathologic finding revealed that lymphangioma circumscriptum. During 11 months follow up, no relapse was found. Conclusion: We described a rare case of lymphangioma circumscriptum on scalp. By using a tissue expander and excision, we achieved no recurrence and aesthetically satisfactory outcome.

Anesthetic Management of the Dental Treatment in a Child with Liver Failure Scheduled for Liver Transplantation - A case report - (간이식 예정인 간부전 환아의 치과치료 시 마취관리 -증례 보고-)

  • Park, Chang-Joo;Jang, Ki-Taeg;Yum, Kwang-Won;Kim, Hyun-Jeong
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.2 no.2 s.3
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    • pp.114-117
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    • 2002
  • Special anesthetic considerations were required for children with acute or chronic liver disease. We experienced a case of dental treatment to control infection under general anesthesia in the 2-year-old girl with liver failure. She was also scheduled for liver tansplantation. Her preanesthetic results of liver function test, electrolytes, and coagulation panel were unstable and out of normal ranges. Uneventful anesthetic induction using isoflurane and atracurium and nasotracheal intubation were carried out. General anesthesia was maintained with isoflurane for 2 hours. Oozing from multiple extraction sites was sustained, so the transfusion of platelet concentration 1 units, fresh frozen plasma 1 unit, and packed red blood cell 1 unit was done. She was recovered without complication but was transferred to pediatric intensive care unit for wound care with her endotracheal tube kept. She was transferred to a ward without noticeable complications next day. So we report this successful case of anesthetic management for dental treatment in a child with liver failure.

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A Study on the Thermal Analysis for the Robotic Arm of the Cord Blood Storage Tank (제대혈 용기 내부 로봇 암의 열해석에 관한 연구)

  • Yun, Sang-Kook;You, Sam-Sang
    • Journal of Advanced Marine Engineering and Technology
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    • v.32 no.5
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    • pp.724-729
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    • 2008
  • Umbilical cord blood has been recently considered an attractive potential alternative as a source of stem cell transplantation to curing diseases such as leukemia, cancers, immune disorders. Normally the stored system of the umbilical cord blood specimen is equipped with a computer-controlled robotic arm that enables the samples to locate the identification places in liquid nitrogen tank at regulated temperature as about $-196^{\circ}C$. As the half of robotic arm is in the air and the rest part is submerged in liquid nitrogen, the temperature of robotic arm varies from ambient to liquid nitrogen temperature. In this study the temperature variation of upper part of arm above tank lid was thermally analysed by using the commercial code of Ansys. The result of analysis was that the upper part of robotic arm was seriously frozen due to heat transfer from liquid nitrogen as low as -$120^{\circ}C$. In order to solve the frost problem of robotic arm, small PTFE tube block as resistance material was introduced into the lower part of tank lid instead of the whole stainless steel(SUS) robotic arm. The results showed that the temperature of robotic arm above the lid was higher enough, and this method would be one of the very effective measure to solve the problem.

Immunohistochemistry for detection of Aujeszky's disease virus antigens : Immunogold-silver method in tissue sections (오제스키병 바이러스 항원검출을 위한 면역조직화학적 연구 -조직절편내 immunogold-silver법-)

  • Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.365-369
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    • 1988
  • The present study was done to demonstrate ADV antigens in frozen and paraffin sections from ADV-infected pigs and cell cultures by using of the IGS method. Tissue specimens from 3 young pigs infected with ADV-phylaxia strain and of 2 healthy pigs were used. Fibroblastic cells originated from pig brain and BHK cells were grown and confluent monolayers were infected with the virus. Two monoclonal antibodies and a specific hyperimmune serum to ADV were used as the source of primary antibodies for both the IGS and immunoperoxidase methods. Application of the IGS method yielded a black fine granular reaction in positive areas, and the results were superior to those obtained using the immunoperoxidase technique for all cases tested. The IGS method might be useful in the detection of various viral antigens in tissue sections.

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A Case of Osteosarcoma of the Mandible (좌측 하악골에 생긴 골육종(Osteosarcoma) 1예)

  • Kim Chan-Jong;Lee Jae-Dong;Nam Soon-Yuhl
    • Korean Journal of Head & Neck Oncology
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    • v.19 no.2
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    • pp.179-183
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    • 2003
  • Osteosarcoma is one of the most frequently occurring malignant bone tumor except for multiple myeloma. However, osteosarcoma of the mandible is rare and aggressive malignancy constituting about 5% to 13% of all cases of skeletal osteosarcoma. The authors experienced a case of osteosarcoma of the mandible in a 31-years old male patient. He visited our outpatient clinic with painless mass of the cheek. On the basis of the clinical and imaging findings, the differential diagnosis included giant cell tumor, histiocytosis and malignant bone tumor. In the first operation, the mass of the mandible was excised. Intraoperative microscopic examination of a frozen section did not yield a diagnosis: however, the final definitive histologic examination revealed osteoblastic osteosarcoma. After 15days, wide excision with hemimandibulectomy was completed followed by postoperative radiotherapy and chemotherapy. With our experience, we report the progression of diagnosis and management for mandibular osteosrcoma.