초기 임신우의 공란우 활용이 초음파 유도 난자 채취 및 수정란 생산에 미치는 영향

진종인,권태현,최병현,김성수,조현태,방재일,김삼철,조규완,이정규 韓國受精卵移植學會 2011 한국동물생명공학회지 Vol.26 No.1

본 연구에서는 유전적 가치가 높은 가축을 OPU 기술을 이용하여 단기간에 반복적이고 연속적으로 생체 내 난포란을 채란하여 수정란을 생산할 수 있는 가능성을 연구한 것으로써, 수정란의 안정적인 생산 및 이식으로 가축 개량의 세대 간격을 단축하기 위해 우수한 유전력을 가진 염선된 개체의 임신 초기 3개월 동안에 반복적인 수정란 생산 가능 여부를 조사하였다. 1. 비임신우 및 임신 초기우에 2회/주 채란으로 비인신우에서는 68회 채란으로 생성된 난포수는 72 This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were and , and per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was and (MeanSD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then and blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.

OPU(Ovum Pick-Up) 채란기간이 난자 및 수정란 생산에 미치는 영향

진종인,권태현,최병현,김성수,조현태,공일근 韓國受精卵移植學會 2010 한국동물생명공학회지 Vol.25 No.1

본 연구에서는 OPU를 통한 체내 유래 난자를 이용한 수정란 생산 시 1개월에서 6개월까지의 연구 기간에 따른 난포 생성수, 난자 회수율, 난자 등급율, 배반포 생성률을 분석하여 공란우의 활용 기간에 관하여 조사하였다. 1. 채란 기간에 따른 난포 생성수는 1개월에서 5개월까지의 난포 생성수는 , , , , 개로 유의적인 차이가 없었으나, 6개월째에는개로 급격하게 줄어든 것을 알 수 있었다. 2. 채란 기간에 따른 난자 회수개수는 1개월에서 3개월까지는 This study was performed to identify the optimal timing for oocyte donor replacement during OPU procedure. OPU was carried out to collect oocytes from every donor at an interval of days (2 times a week). The collected oocytes were matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml of FSH and 1 mg/ml of estradiol for 24 h. After 24 h of exposure to sperm, the presumptive zygotes were cultured in CR1aa medium supplemented with 4 mg/ml of BSA for 3 days before being changed to CR1aa medium with 10% of FBS for another days. The mean numbers of retrieved oocytes were remained constantly up to 3 months (, , ), but significantly decreased at over 4 to 6 months (, , ) (p<0.05). The blastocyst development potential was also very similar rate from 1 to 3 months (37.2%, 40.4% and 44.6%), but significantly decreased from 4 to 6 months (24.8%, 29.3% and 28.6%, respectively) (p<0.05). The production of OPU derived embryos in periods of 1 to 3 months (, and ) were significantly higher than those in 4 to 6 months (, and , respectively) (p<0.05). In conclusion, the efficient periods for the production of OPU derived embryos was until 4 months, twice per week to produce over 64 transferable embryos and then replace new donor after 3 months use. The best replacement time is 3 months and could be maximized production of OPU derived embryos.

Application of Artificial Insemination Technology for Dairy Breeding in Mongolia

진종인,김성수,조현태,최병현,이정규,김윤식,김삼철,조규완,Tumor Baldan,공일근 사단법인 한국동물생명공학회 2011 한국동물생명공학회지 Vol.26 No.4

This study was focused on improvement of milk production in Mongolian dairy industry by artificial insemination (AI)technology, supported by ODA of KOICA in Republic of Korea. This program was started in January 2009 and it is in 3rd years. This manuscript summarized the data especially on estrus synchronization and pregnancy establishment in dairy cows (Holstein) this year. A total of 81 dairy cows from 4 private farms (38 from Undarmal milk and that of 30, 8 and 5 dairy cows from Onjin (Enkhbayer), Jargalant, and BRM School farms respectively) were synchronized with 5 ml Lutalyse (i.m.) in the dump of dairy cows and then estrus was detected 2 to 3 days after PGF2α injection. The synchronized dairy cows were inseminated with 0.5 ml dairy frozen semen by conventional artificial insemination (AI) techniques. Pregnancy was diagnosed about 60 days after AI by palpation method. About 96.3% (78/81) of synchronized cows were responded to single PGF2α injection. Total 75 over 78 dairy cows (90.1%) inseminated were diagnosed as pregnant. The estrus induction and pregnancy rates were very effective using Lutalyse injection and conventional AI techniques in Mongolian dairy cow. The present results indicated that AI after estrus induction in Mongolian dairy cows could be applied to dairy breeding technology for improving breeding efficiency and milk production of the country

OPU 유래 한우 수정란 생산 및 이식

진종인,최병현,김성수,조현태,선두원,임현태,이정규,민찬식,공일근 사단법인 한국동물생명공학회 2014 한국동물생명공학회지 Vol.29 No.3

This study was carried out to establish the system of OPU derived embryo production, management of recipientsas well as offspring production. OPU derived embryo production system was carried out of aspiration of immatureoocytes 2 times per week, total 24 times for 3 months by an ultrasonographic guided follicular aspiration system andthen produced in vitro-produced blastocysts by in vitro maturation, fertilization and culture system. This work wascollected total 13,866 oocytes, average 8.2 ± 4.5 oocytes per session and 8,170 G1 + G2 grade oocytes, average 4.8oocytes per session by 1,692 times session of total 71 donors for 4 years from 2010 to 2013. The rate of cleavageand blastocyst developmental competence were obtained 11,825 (85.3%) and 5,032 (36.3%) that was 7.0 ± 3.8 cleavedembryos and 3.0 ± 2.5 blastocysts per session. OPU derived embryo transfer were taken place in 2, 4, 6 and 7 localgovernments at 2010, 2011, 2012 and 2013 for 4 years and pregnancy rate were obtained 41.2, 43.9, 46.5 and 49.7%in each years. It means that pregnancy rate was continuously improved according of every year for 4 years. Pregnancyrate was significantly different according to individual local government in which was 62.7% in B, but 24.2% in Fat 2012. Paternity identification was carried out total 26 offspring in C local government of 2012 and then confirmed100% agreement of its analysis. In conclusion, the results obtained the possibility of mass production of elite cowembryos as well as offspring by OPU derived embryo production system, of which could be decreased the requiredtime of genetic improvement.

젖소에서 초음파기기를 이용한 난자 채취에 있어서 손가락 촉지를 이용한 난포란의 채란

진종인,홍승표,정장용,이지삼,박희성 韓國受精卵移植學會 2000 한국동물생명공학회지 Vol.15 No.3

This study was conducted to develop an improved method for oocyte pick-up(OPU) with finger-sensibility using ultrasound-guidance from ovarian follicles in Holstein heifers. Oocytes were aspirated from ovarian follicles of clear-outline (>2mm), obscure-outline and invisible( 2mm) on ultrasound images with 3 different vacuum pressure(40, 80, 120mmHg). Total number of oocytes recovered/follicles were 309/237(130.4%). 113/80(141.3%) and 107/74(144.6%) with 40, 80 and 120 mmHg of vacuum pressure, respectively. Mean number of oocytes recovered was higher in 2 OPU/week (18.35.3) than 1 OPU/week(14.54.1), but this difference was not statistical1y significant. The recovery rates were not affected by the number of OPU as 135.6%(282 oocytes/208 follicles) in 1~20 OPU, 137.7% (168/122) in 21~40 OPU and 148.4%(92/62) in 41~60 OPU, respectively. The proportions of good oocytes (Grades I) recovered were not significantly different by the number of OPU until 40 OPU(12.4% in 1~20 OPU vs 16.7% in 21~40 OPU). However, a significantly(P<0.05) lower recovery rate resulted from more than 40 OPU compared to less than 40 OPU(7.6%). These results imply that more fertilizable oocytes can be produced from invisible-immature follicles by transvaginal aspiration with finger-sensibility from Holstein heifers.

OPU 유래 수정란의 대량생산을 위한 고능력 공란우 반복사용 가능성에 관한 연구

진종인,최병현,김성수,박번영,이정규,공일근 사단법인 한국동물생명공학회 2015 한국동물생명공학회지 Vol.30 No.3

This study was designed to know the possibility in repeat uses of elite donor cows for getting mass production of OPU-derived embryo production (OPU-IVP). Ultrasound transvaginal ovum pick-up (OPU) performed in 6 Korean native cows was aged 4 to 10 years old. The aspiration of immature oocytes for OPU derived embryo was carried out 2 times per week, and OPU-IVP of 1st period was carried out 22∼48 sessions from each donors. And the break time for OPU-IVP of 2nd period after 1st OPU from each donors were 2∼25 months. The OPU-IVP of 2nd period each donors conducted total 15∼65 times for 2∼8 months by an ultrasonographic, was guided follicular aspiration system. The average numbers of collected oocytes, grade 1 + grade 2(G1+G2) oocytes and cleavage embryo from 1st period OPU-IVP were significantly differences between donors (p<0.05). Total collected oocytes of donor D were significantly higher compared with donors of A, B, C, E and F (average 17.0 per session vs. 11.2, 10.1, 8.5, 10.2 and 9.6; p<0.05) and also oocytes of G1+G2 were significantly higher compared with r A and D and subsequently to donors of B, C, E and F (average 7.9 and 8.5 per session vs. 5.0, 2.7, 6.0 and 1.6; p<0.05). Cleavage rate of donor D was significantly higher compared with donors of A, B, C, E and F (average 13.1 per session vs. 10.1, 9.1, 6.9, 8.9 and 6.7; p<0.05). The average numbers of OPU-IVP for 1st period was significantly higher from donors of B, D and E than those from donors of A, C and F (average 6.5, 7.1 and 6.5 per session vs. 3.5, 4.2 and 2.8; p<0.05). The possibility investigation of 2nd OPU-IVP was carried out after 2∼25 months rest periods from 1st period OPU session. Total average numbers of collected oocytes, cleavages and blastocyst development rates were significantly higher from 1st period OPU compared with 2nd period one (p<0.05). The OPU-IVP efficiency by break for more embryo production from elite cow was analysis comparing without rest of donor A, under 6 months rest period as B and over 6 months rest period as C and then the average numbers of collected oocytes, cleavages and blastocysts were significantly higher from A group (11.8, 9.5 and 5.2 per session) than those from B and C groups (7.9, 6.2 and 2.6 vs. 9.2, 7.5 and 3.9, p<0.05), and also C group was significantly higher than B group. In conclusion, 1st period OPU-IVP was more efficient compared with 2nd period repeated uses of donor, and the break times for additional production of embryo on donor were needed more than over 6 months after 1st period OPU-IVP. This repeating uses of elite donor cows given more emphasis for getting the opportunity on mass production of elite cow OPU-IVP embryo should be increased G1+G2 possibility of genetic improvement of livestock within short period.

DNA 검정우에서 FSH 호로몬 투여가 난포란의 채란율에 미치는 영향

진종인,이명열,박준규,이지삼,정장용,박희성 한국수정란이식학회 2002 한국수정란이식학회 학술대회 Vol.2002 No.1

초음파기기를 이용한 난포란의 채란 기술은 유전적으로 우수한 수정란의 다량확보가 가능함으로써 가축개량에 널리 응용할 수 있다. 그러나 초음파유도 난포란의 채란은 무엇보다도 회수율과 회수한 난포란의 질적 등급의 개선이 이루어져야 할 것이다. 본 연구는 등지방층두께, 일당증체량, 근내지방도 및 배최장근 단면적에 연관된 DNA marker가 검정된 한우로부터 FSH(FOLLTROPIN-V, Vetrepharm, Canada)를 투여하여 FSH 투여전.후의 포의 발달율과 등급별 회수율을 조사하였다. 공시동물의 발정주기에 관계없이 매주 무처리와 채란 12시간 전에 FSH를 각각 l00mg 과 200mg을 근육 주사하여 난포란의 채취를 유토하였다. 난소내 난포란의 채란은 SONOACE-600형 (Medison CO., 한국) 초음파기기를 사용하여 모니터 상에 검게 나타나는 난포를 17-gauge needle (Cook, Australia) 로써 70mmHg로 유지된 regulated vacuum pump를 이용하여 모두 채란하였으며, 소난포는 손가락 촉지를 이용하여 채란하였다. 난포의 크기는(large : 6mm, medium : 2-6mm, small 2mm) 무처리구에서 small(56.6%), medium(33.0%), large follicles(10.4%) 순으로 나타났으며, 전체 난포수는 106(8.23.6개) 였다. FSH 200mg 투여구에서는 medium(47.5%), small(32.5%), large follicles(20.0%) 순으로 나타났으며, 전체난포수는 40(10.02.2개) 였다. FSH l00mg 투여구에서는 medium(45.6%), small(43.5%), large follicles (10.9%) 순으로 나타났으며, 전체 난포는 92(7.72.5개) 였다. FSH 투여전 초음파 image 상으로 관찰한 난포수와 FSH 투여 후 초음파기기와 손가락 촉지를 이용하여 난포의 발달 및 난포란의 채란율은 200mg 투여 구에서는 투여 전에 4.30.6개였으나 투여 후에는 9.32.1개로 증가하였으며, 채란율은 75.0% 였다. l00mg 투여구에서도 투여 전후에 각각 5.31.2개 및 8.22.3개였으며, 채란율은 81.6%였다. 난포란의 채란율은 무처리구에서 88.7%(7.23.9개)로 나타났으며, 회수한 난포란의 등급은 각각 0.0%(GI), 5.3%(GII), 28.T%(GIII), 66.0%(GIV)로 나타났다. FSH 200mg 투여구의 채란율은 87.5%(8.82.6개)로 나타났으며, 등급별회수율은 GI(2.8%), GII(8.6%), GIII(34.3%), GIV(54.3%)으로 나타났다. 뿐만 아니라 FSH 100mg 투여구에서도 회수율은 89.1%(6.85.2개)였으며, 등급별 회수율은 각각 GI(8.5%), GII(13.4%), GIII(43.9%), GIV(34.2%)로 나타났다.

광섬유 간섭계를 이용한 웨이퍼의 두께 및 굴절률 동시 측정

진종한(J. Jin),맹새롬(S. Maeng),김종안(J.-A. Kim),강주식(C.-S. kang),김재완(J.W. Kim) 한국정밀공학회 2014 한국정밀공학회 학술발표대회 논문집 Vol.2014 No.5월

높은 동적 범위를 갖는 간섭 기반 두께 측정 기술

진종한(J. Jin),박정재(J. Park),배재석(J. Bae) Korean Society for Precision Engineering 2023 한국정밀공학회 학술발표대회 논문집 Vol.2023 No.11

현대 무용수들의 골반과 고관절의 상해에 대한 연구

진종은 고려대학교 대학원 통계학과 1999 통계상담 Vol.14 No.2