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우 난포란의 (卵胞卵) 체외성숙에 관한 연구 Ⅰ. 난포란의 회수 및 체외배양
윤산현(S . H . Yoon),고대환(D . H . Ko),박세필(S . P . Park),박태균(T . G . Park),정길생(K . S . Chung) 한국축산학회 1989 한국축산학회지 Vol.31 No.4
These experiments were carried out to develop the effective recovering method of ovarian follicular oocytes and to investigate the optimum culture conditions for in vitro maturation of recovered follicular oocytes. Bovine follicular oocytes with various configurations of cumulus cell mass were collected by aspirating follicular fluid from the ovarian follicles of various size, and then cultured in various media for 28h in a incubator with 5% C0₂ in air at 38℃. The results obtained in these experiments were summarized as follow: 1. The average number of the follicular oocytes recovered from a cow was 46.4 and the highest recovery rate was obtained from middle size follicles, 3-5㎜ in diameter. 2. The majority of the follicular oocytes with compacted cumulus cells existed in GV stage While those with dispersed cumulus cells or without cumulus cells existed between GVBD and MⅡ stage. 3. The extrusion rate of Ist polar body of the follicular oocytes cultured in Ham`s F-10, m-KRB and BMOC-3 containing hormones was not higher than those of oocytes cultured in those media without hormones. However, in case of m-TALP medium, the highest extrusion rate of Ist polar body, 77.7% was obtained by adding hormones such as FSH, HCG and Estradiol. 4. The maturation rate of the follicular oocytes cultured in basic medium without or with matured cumulus cell was 40% or 51.1%, respectively, with significant difference(p$lt;0.05).
백청순,한용만,이경광,정길생,김종배,고대환 ( C . S . Baik,Y . M . Han,K . K . Lee,K . K . Chung,J . B . Kim,D . W . Ko ) 한국축산학회 1986 한국축산학회지 Vol.28 No.11
These experiments were carried out to obtain basic information necessary to produce the offsprings which were sexed by chromosomal analysis prior to transfer in the mouse. Morula stage embryos were obtained at 3 clays post-coitum. Embryos were mechanically bisected by a glass microneedle and then the pairs of demi-embryos were cultured in BMOC-3 medium under the humidified atmosphere of 5% CO₂ at 37℃ for 22-26 hours. To increase the appearance of metaphase, bisected embryos normally developed to the blastocyst were cultured in MOC-3 medium containing colcemid (0.04㎍/㎖) under the same condition for 3 hours. One of both demi-embryos was sexed by chromosomal analysis and the other was transferred to the uterine horn of a foster mother. The results obtained in these experiments were summarized as follows; 1. Of 241 pairs of demi-embryos cultured for 22-26 hours, 162 pairs (67.2%) were normally developed to the blastocysts. In the control groups, the percentage of intact and zona-free embryos normally developed to the blastocysts was 96.5% and 93.9%, respectively. 2. The mean Mitotic Index of 121 demi-embryos subjected to chromosomal analysis was 11.3±13.2%r and 18 demi-embryos (14.9%) were sexed. 3. Of 18 demi-embryos transferred to recipients after sexing, 2 offsprings were born and their sex was corresponded to the sex determined by chromosomal analysis.
X- 정자와 Y- 정자의 분리에 관한 연구 2 . Percoll 중층원심분리법에 의한 인간정자의 분리
엄기붕(K . B . Oum),이주영(J . Y . Lee),고대환(D . H . Ko),정길생(K . S . Chung) 한국축산학회 1988 한국축산학회지 Vol.30 No.5
These experiments were carried out to develop new techniques for in vitro separation of X-and Y-bearing spermatozoa. One ㎖ of washed human sperm suspension was loaded on the isotonic discontinuous Percoll density gradient, and then it was centrifuged at 250×G for 25 min. After centrifugation, spermatozoa were fractionated according to Percoll density gradient. Spermatozoa included in each fraction were subjected to the estimation of motility, morphological abnormality, F-body test, and recovery rate of spermatozoa was also investigated. The results obtained in these experiments were summarized as follows: 1. Following centrifugation of discontinuous Percoll density gradient, population of spermatozoa increased progressively from low density to high density. The highest concentration of spermatozoa was observed in 7th fraction which included 20% of spermatozoa. 2. High percentage of motile spermatozoa was observed at high Percoll concentration and the highest percentage was obtained at 6th fraction. 3. Following Percoll centrifugation, percentage of X-sperm increased from 53.2% (control) to 74.1% (7th fraction). 4. Following centrifugation, sperm abnormality was increased at low Percoll gradient and decreased at high Percoll gradient. The lowest abnormality (15.0%) and the highest abnormality (46.0%) were observed at 7th fraction and 1st fraction, respectively.