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사람 난포액의 처리 방법과 Sample이 생쥐 수정란의 체외 발달율에 미치는 영향
전병균,최연희,조은정,송건호,곽대오,문진수,김광철,Jeon, Byeong-Gyun,Choi, Yeon-Hee,Jo, Eun-Jung,Song, Gun-Ho,Kwak, Dae-Oh,Moon, Jin-Soo,Kim, Kwang-Chul 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.4
The present studt was performed to investigate the effect of treatment and samples of human follicular fluid (hFF) on the development in vitro of mouse embryos. The two cell stage embryos collected at 40 h post-hCG injection were cultured in the modified human tubal fluid (m-RTF) containing 15% synthetic serum substitute (SSS) or human tubal fluid (hFF) for up to 3 days at $37^{\circ}C$ in 5% $CO_2$ incubator. Also the composition of hormone, total protein and protein pattern of hFF samples were analyzed. The developmental rate of mouse embryos developed to blastocyst were not significant difference in the m-RTF containing 15% hFF filtered with 0.22 or 0.8 ${\mu}m$ syringe filter, however, the embryos cultured in the m-RTF containing inactivated hFF were significantly (p<0.05) developed at the high rate to blastocyst than those containing fresh hFF and SSS. The in vitro developmental rate to blastocyst and hatched blastocyst in the m-RTF containing 15% hFF sample A (90.5 and 85.4%, respectively) and SSS (79.4 and 75.3, respectively) were significantly (p<0.05) increased, compared with hFF sample B (64.2 and 54.1 %, respectively). The hFF sample A tended to be higher concentration of LH, FSR, total protein and the ratio of progesterone/$E_2$ and lower concentration of $E_2$ and progesterone than the hFF sample B, but there were no differences in the protein pattern between the two hFF samples. The results of these study suggest that the addition of hFF to the culture medium enhances the development in vitro to blastocyst and hatched blastocyst, but the in vitro developmental rate of mouse embryos is different between hFF samples.
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세포질내 정자주입법(ICSI)에 있어서 정자흡입 및 난자내 주입방법에 관한 연구
이택후 ( Taek Hoo Lee ),김항진 ( Hang Jin Kim ),송건호 ( Gun Ho Song ),김대근 ( Dae Geun Kim ),전상식 ( Sang Sik Chun ),박윤규 ( Yoon Kyu Park ),서태광 ( Tae Kwang Suh ),전병균 ( Byeong Gyun Jeon ),류은경 ( Eun Kyung Ryu ),이은숙 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.12
Immobilization of spermatozoa prior to intracytoplasmic sperm injection(ICSI) sometimes results in crooked tail and this makes it difficult to aspirate sperm into an injection pipette tail first Head-first sperm aspiration into an injection pipette avoid this problem due to the bigger size of the sperm head. The effect of head or tail-first sperm injection into an oocyte on fertilization, cleavage, percentage of grade 1 embryos and development to blastocyst stage in ICSI program has been studied. A single living immobilized spermatozoa from oligoasthenozoospermic patient was injected into an oocyte head-first or tail-first according to the treatment. Eighteen hours after microinjection, oocytes were inspected for survival and fertilization Fertilized oocytes with two pronuclei were cultured in 30 μ I drop of mHTF supplemented with 10 % heat-inactivated follicular fluid(FF) at 37℃. On day 2, embryo transfer was performed with cleaved embryos. The remaining 2-8 cell stage embryos were co-cultured with BRL cells in mHTF+10 % FF for 72 hours and the developmental stage was observed. The data were analyzed by Analysis of Variance. A total of 164 oocytes from 36 cycles were assigned to each treatment and ICSI was performed(88 head-first, 76 tail-first). The rates of normal fertilization were 81.8 % and 76.3% for head-first and tail-first, respectively. Of the fertilized oocytes, the percentage of cleaved embryos and the percentage of grade 1 embryo among cleaved embryos were 88.9 % and 68.8 %, 93.1 % and 74.1 % for head-first and tail-first, respectively. Of the 2-8 cell embryos cultured, 44.4 %(16/36) and 50.0%(10/20) for head first and tail first, respectively developed to blastocyst stage. There were no differences in fertilization, cleavage, rates of grade 1 embryos, and development to blastocyst stage. In conclusion, head-first or tail-first sperm injection into an oocyte in ICSI program does not affect fertilization and subsequent embryo development to blastocyst stage in vitro.
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세포질내 정자주입법(ICSI)에 있어서 정자흡입 및 난자내 주입방법에 관한 연구
이택후,김항진,송건호,김대근,전상식,박윤규,서태광,전병균,류은경,이은숙,문진수,김광철 경북대학교 의학연구소 2000 경북대학교병원의학연구소논문집 Vol.4 No.1
Study on Method of Sperm Aspiration and Injection into an Oocyte in Intracytoplasmic Sperm Injection(ICSI) Immobilization of spermatozoa prior to intracytoplasmic sperm iniection(ICSI) sometimes results in crooked tail and this makes it difficult to aspirate sperm into an injection pipette tail first. Head-first sperm aspiration into an injection pipette avoid this problem due to the bigger size of the sperm head. The effect of head or tail-first sperm injection into an oocyte on fertilization cleavage, percentage of grade I embryos and development to blastocyst stage in ICSI program has been studied. A single living immobilized spermatozoa from oligoasthenozoospermic patient was injected into an oocyte head-first or tail-first according to the treatment. Eighteen hours after microinjection, oocytes ware inspected for survival and fertilization Fertilized oocytes with two pronuclei were cultured in 30μl drop of mHTF supplemented with 10% heat-inactivated follicular fluid(FF) at 37℃. On day 2. embryo transfer was performed with cleaved embryos. The remaining 2-8 cell stage embryos were co-cultured with BRL cells in mHTF + 10% FF for 72 hours and the developmental stage was observed. The data were analyzed by Analysis of Variance. A total of 164 oocytes from 36 cycles were assigned to earth treatment and ICSI was performed(88 head-first, tail-first). The rates of normal fertilization were 81.8% and 76.3% for head-first and tail-first, respectively. Of the fertilized oocytes, the percentage of cleaved embryos and the percentage of grade 1 embryo among cleaved embryos were 88.9% and 68.8%, 93.1% and 74.1% for head-first and tail-first, respectively. Of the 2-8 cell embryos cultured, 44.4%(16/36) and 50.0%(10/20) for head first and tail first, respectively developed to blastocyst stage. There were no differences in fertilization, cleavage, rates of grade 1 embryos, and development to blastocyst stage. In conclusion, head-first or tail-first sperm injection into an oocyte in ICSI program does not affect fertilization and subsequent embryo development to blastocyst stage in vitro.
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