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동결보존시 생쥐 전핵배아의 시기에 따른 생존율과 발생율의 비교
김희선,류범용,오선경,서창석,김석현,최영민,김정구,문신용,이진용,Kim, H.S.,Ryu, B.Y.,Oh, S.K.,Suh, C.S.,Kim, S.H.,Choi, Y.M.,Kim, J.G.,Moon, S.Y.,Lee, J.Y. 대한생식의학회 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.1
This study was designed to evaluate the influence of pronuclear age on the survival and post-thawing development after cryopreservation of mouse embryos. Freezing and thawing were performed in the different pronuclear stages of mouse embryos after IVF. Embryos were obtained from $F_1$ hybrid mice and classified into 4 groups according to the pronuclear stage (6hr, 9hr, 12hr and 15hr after insemination). Pronuclear ova were slowly cooled in a biological freezer using 1.5M 1,2-propanediol and 0.1M sucrose as cryoprotectant. Thawing was done at room temperature and 1,2-propanediol was removed by multi-step dilutions. Both frozen-thawed embryos and control fresh embryos were cultured in vitro in Ham's F-10 medium supplemented with 4mg/ml BSA. In control group, the development rate after 48hr was 99.3%, and the complete hatching rate after 144hr was 61.3%. In experimental groups, the survival rate after thawing was 95.4% in 6hr, 88.7% in 9hr, 75.2% in 12hr and 62.4% in 15hr after insemination, the development rate after 48hr was 61.1, 77.0, 67.0 and 79.6%, respectively, and the complete hatching rate after 144hr was 25.7, 43.7, 42.2 and 60.0%, respectively. The survival rate in 15hr was significantly lower (p<0.05) compared with other groups. In vitro development rates after 48hr were similar in all groups, but complement hatching rate was significantly lower (p<0.05) in 6hr group. In conclusion, cryopreservation of mouse pronuclear ova with 2 distinct pronuclei (9hr and 12hr groups) showed better results after thawing compared with early (6hr group) or late pronuclear ova just prior to cleavage (15hr group).
김희선,설혜원,안희진,오선경,구승엽,김석현,최영민,김정구,문신용,Kim, Hee-Sun,Seol, Hye-Won,Ahn, Hee-Jin,Oh, Sun-Kyung,Ku, Seung-Yup,Kim, Seok-Hyun,Choi, Young-Min,Kim, Jung-Gu,Moon, Shin-Yong 대한생식의학회 2004 Clinical and Experimental Reproductive Medicine Vol.31 No.4
Objective: This study was performed to evaluate the possibility of prolonged culture of human embryonic stem cells (hESC; SNUhES2) on human amniotic fluid cells (hAFC), which had been storaged after karyotyping. Method: The hAFC was prepared for feeder layer in the presence of Chang's medium and STO medium (90% DMEM, 10% FBS) at $37^{circ}C$ in a 5% $CO_2$ in air atmosphere. Prior to use as a feeder layer, hAFC was mitotically inactivated by mitomycin C. The hESCs on hAFC were passaged mechanically every seven days with ES culture medium (80% DMEM/F12, 20% SR, bFGF). Results: The hAFC feeder layer support the growth of undifferentiated state of SNUhES2 for at least 59 passages thus far. SNUhES2 colonies on hAFC feeder appeared slightly angular and flatter shape as compared with circular and thicker colonies observed with STO feeder layer and showed higher level with complete undifferentiation in seven days. Like hESC cultured on STO feeders, SNUhES2 grown on hAFC expressed normal karyotype, positive for alkaline phosphatase activity, high telomerase activity, Oct-4, SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 and formed embryoid bodies (EBs). Conclusion: The hAFC supports undifferentiated growth of hESC. Therefore, these results may help to provide a clinically practicable method for expansion of hESC for cell therapies.
국내 야생따쥐(Crocidura lasiura) 의 Hantavirus 항체 보유실태 및 원인체 분리
김희선,강문일,Kim, Hee-sun,Kang, Mun-il 대한수의학회 1994 大韓獸醫學會誌 Vol.34 No.1
Eleven shrews were caught from three areas of Korea. All of them were confirmed in the same species, Crocidura lasiura. All of sera from wild shrews were examined by indirect immunofluorescent test against Hantaan-related virus. The antibody to Hantaan-related virus was detected by 2 out of 11 shrews. Just 2 of 7 shrews from BG area were sero-positive for Hantaan-related virus antigen and none from other. All of sero-positive for Hantaan-related virus antigen belonged to male with antibody titer of 1:40 to 1:80. Two Hantaan-related viruses were isolated in vivo and in vitro.
국내 야생 집쥐(Rattus norvegicus)의 murine typhus 감염조사
김희선,강문일,고홍범,Kim, Hee-sun,Graham, Robert Ross,Kang, Mun-il,Koh, Hong-bum 대한수의학회 1994 大韓獸醫學會誌 Vol.34 No.4
Sixty eight wild rats, Rattus norvegicus, were caught from Seoul city, Kyonggi, Kangwon, Cholla, and Kyongsang-provinces. All sera collected from rats were examined by immunofluorescent test to find antibody against murine typhus. The antibody prevalance to murine typhus was 4.4%(3/68). Sero-positive rate was 12.5%(2/16) in Kyonggi province and 12.5%(1/8) in Cholla province. The sero-positive rate difference between sexes of rats was not recognized. Three Rattus norvegicus having antibody to murine typhus were morphologically classified as subadult with 8.3%(1/12), middle-aged adult with 5.9%(1/17), and old adult with 4.8%(1/21), respectively.
국내 야생 집쥐류(Rattus norvegicus와 R rattus)의 spotted fever group rickettsia 감염에 관한 역학조사
김희선,강문일,Kim, Hee-sun,Kang, Mun-il 대한수의학회 1995 大韓獸醫學會誌 Vol.35 No.3
Seventy wild rats were caught from Seoul city, Kyonggi, Kangwon, Honam, and yongnam provinces. All of them were in same species, Rattus norvegicus, except two R rattus from Kyonggi province. Seventy sera from wild rats were studied by immunofluorescent antibody assay for evidence of infection by spotted fever group rickettsia. The antidody prevalance was 37.14%(26/70) for spotted fever group rickettsia. The sero-prevalence rates for spotted fever group rickettsia antibody was the hightest in Kyonggi province with 55.56%(10/18), yongnam province with 50.00%(10/20), Kangwon province with 25.00%(2/8), Seoul city with 18.75%(3/16), and Honam province with 12.50%(1/8). The sero-positive rates difference between sexes were higher in female with 46.15%(12/26) than in male with 31.81%(14/44) for spotted fever group rickettsia. Twenty six of 68 Rattus norvegicus with antibody for spotted fever group rickettsia were in subadult with 50.00%(6/12), young adult with 38.89%(7/18), middle-aged adult with 35.29%(6/17), and old adult with 33.33%(7/21). No antibody was detected from R rattus.
조직내 효소형 분석에 의한 집쥐류의 분류학적 특성 조사
김희선,강문일,이재일,Kim, Hee-sun,Kang, Mun-il,Lee, Jae-il 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.4
Sixty wild rate (fifty eight of Rattus norvegicus and two R rattus) were caught from Seoul, Kyonggi, Kangwon, Honam, and Yongnam areas between August and October 1992. From liver homogenates of the wild rats, isoenzyme patterns were analysed by starch gel electrophoresis. Using 9 enzyme systems, eight electrophoretic types were identified among wild rats with genetic diversity per locus between 0.00 and 0.49 (Mean 0.15). R rattus from Kyonggi (Kanghwa) and Kangwon (Cholwon) were distinct from R norvegicus from nine regions with 0.581 in genetic divergence. Therefore genetic divergence was different not only in interspecies(0.581) but also in intraspecies(0.111~0.375). These data suggested that isoenzyme electrophoresis could be used as a potential application in taxonomic studies of wild rats.
국내 야생 집쥐(Rattus norvigicus)의 유행성 출혈열바이러스 감염증의 역학조사
김희선,이준학,강문일,고홍범,Kim, Hee-sun,Graham, Robert Ross,Lee, Jun-hak,Kang, Mun-il,Koh, Hong-bum 대한수의학회 1993 大韓獸醫學會誌 Vol.33 No.2
Sixty eight wild rats were caught from Seoul, Kyonggi, Kangwon, Cholla, and Kyongsang provinces in Korea. All of them were confirmed in the same species, Rattus norvigicus. All of sera from wild rats were examined by indirect immunofluorescent test against Hantaan-related virus. Detection of anti-body to Hantaan-related virus were 4 out of 68 rats(5.9%), including 2 of 8(25.0%) rats in Kangwon and 2 of 20 rats(10.0%) in Kyongsang province. No difference of the sero-positive rates between sex of rats tested to Hantaan-related virus was recognized. According to age, four rats with antibodies(1:320~640) to Hantaan-related virus belonged one subadult, one young adult, one middle aged adult, and one old adult, respectively.