The cryopreservation of Hanwoo embryos has become an integral part of assisted reproduction in animal. The objective of this study was to assess the effect of The objectives of this study were: (1) to evaluate the influence of bovine embryo developmental stage on in vitro embryo development after freezing, (2) to study the efficiency compared with conventional freezed embryos at different embryo source. For conventional slow-freezing, day 7 or 8 expanded blastocysts were collected. The standard freezing medium was 1.8 M ethylene glycol (EG). Embryos were equilibrated in 1.8 Methylene glycol(EG) with 0.1 M sucrose in Dulbecco's phosphate-buffered saline (D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25 ml-straw and placed directly into cooling chamber of programmable freezer precooled to , after 2 min, the straw was seeded, maintained at for 8 min, and then cooled to at /min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 see and exposed to water for 20 sec. Straws were then removed from water. Rates of blastocyst survive and hatched were evaluated at 12 to 48h post-warming. The re-expansion and hatched rates of morula embryos were significantly lower than those obtained for blastocysts and expansion blastocysts (31.6%, 10.5% vs, 68.9%, 22.2% vs, 73.7%, 53.6%, respectively). No differences in re-expansion rates were found between in vivo and in vitro blastocysts. whereas hatched rates was significantly higher (51.2%) in vivo compared with in vitro embryos (18.6%). in conclusion, demonstrate that conventional freezing can be used successfully in cryopreservation of in vitro and in vivo bovine embryos, and that it might be considered for use in commercial programs and embryo preservation.
This study was carried out to investigate inflammation-related gene expression altered in ovary and endometrium of Korean cattle with reproductive disorders using microarray. In the present study, nine inflammation-related differential1y expressed genes (DEGs) were identified in the cystic ovary and endometrium with endometritis. In the follicular cyst, eotaxin and alpha-2-HS-glycoprotein (AHSG) were up-regulated, whereas complement component 3 (C3) and oxidised low density lipoprotein (lectin-like) receptor 1 (OLR1) were down-regulated. Complement component 4A (C4A) was up-regulated in luteal cyst. In the endometritis, chemokine 1igand l and 2 (CXCL1 and CXCL2), protein C (inactivator of coagulation factors Va and VIIIa), and complement component C5 were up-regulated, whereas kininogen was down-regulated. Of these genes, we focused on eotaxin and kininogen, which were highly regulated in the follicular cyst and endometritis, respectively and on C3 commonly regulated in both reproductive disorders. The microarray data of eotaxin, kininogen, and C3 were validated by semi-quantitative PCR. Consistent with microarray data, eotaxin was up-regulated by 4-fold in the follicular cyst, while kininogen was down-regulated by 5-fold in the endometritis. C3 was down-regulated in the both follicular cyst and endometritis. Our results suggest that these inflammation-related genes could be useful markers for diagnosis of cystic ovary and endometritis of Korean cattle.
The objective of this study is to investigate the changes in concentrations of leptin and insulin in serum of Korean cattle (Hanwoo) with reproductive disorders and to examine the relationship among leptin, insulin, and body condition score (BCS). The concentration of leptin in serum of pregnant Hanwoo showed insignificant difference from that in serum of Hanwoo with reproductive disorder, such as repeat breeding, follicular cyst, corpus luteum cyst, ovarian atrophy, and feeble estrus (p>0.05). However, the concentrations of leptin and insulin in serum were changed with different BCS value. In emaciated Hanwoo (BCS ), they were significantly decreased compared to BCS (p<0.05). The leptin showed different genotypes with different BCS value. In BCS , C/T genotype was expressed (83.3%) more than C/C (16.7%) or T/T (0%) genotype, whereas C/C genotype was expressed (62.5%) more than C/T (25.0%) or T/T (12.5%) genotype in BCS . The insulin concentration in follicular fluid obtained from ovary with follicular cyst which has follicles having diameter of was significantly higher (p<0.05) than those in normal follicle fluid which has follicles having diameter of . These results showed that concentration of leptin and insulin in serum were related to BCS value and follicular size and suggest that the changes in concentration of leptin and/or insulin in serum could be a potent biomarker for diagnosis of bovine reproductive disorder.
The objective of this study is to investigate the changes in concentrations of blood urea nitrogen (BUN) and sex steroid hormones, such as estrogen, progesterone, and testosterone in Korean cattle (Hanwoo) with reproductive disorders and to examine the relationship between BUN and body condition score (BCS) in Hanwoo. The concentration of BUN was 16.2 mg/dl, 17.8 mg/dl, 15.1 mg/dl, 17.9 mg/dl, and 28.3mg/dl in pregnancy, repeat breeding, follicular cyst, luteal cyst, and ovarian atrophy, respectively. In Hanwoo with BCS , and , the concentration of BUN was 15.8 mg/dl, 17.0 mg/dl, and 17.6 mg/dl, respectively. Fluoroimmunoassay showed that serum estrogen and progesterone levels were decreased in reproductive disorders Hanwoo, such as ovarian atrophy, endometritis, and weak estrus. The testosterone level was significantly decreased in Hanwoo with reproductive disorders compared to that in pregnant Hanwoo ( vs 0.13 ng/ml, p<0.05). The progesterone and estrogen concentrations in follicular fluid obtained from ovary with follicular cyst were significantly higher (p<0.05) than those in normal follicle fluid. These results show that there is no relationship between BUN and BCS in Hanwoo, and the concentration of sex steroid hormone in serum and follicular fluid are changed in reproductive disorders Hanwoo.
본 연구는 돼지 체외수정란 생산효율을 향상시켜 돼지의 품종개량, 형질전환 돼지생산 등과 멸실위험에 처해 있는 유전자원의 보존을 위한 기술로 활용하기 위해 미성숙 난포란의 적정 체외성숙 시간을 알아보고, 배양액의 종류 및 체외 배양시의 산소 농도에 따른 체외수정란의 생산 효율을 확인한 결과는 다음과 같다. 1. 돼지 미성숙 난포란의 체외성숙 시간별 제2감수분열중기(M II)까지 성숙된 비율이 체외성숙 38, 40, 42시간째에 각각 61.1%, 42.9%
임신의 성립 및 유지에 중요한 역할을 하는 자궁내막세포에 통로의 존재를 확인하기 위하여 본 연구를 수행하였다. 통로는 일반적으로 중추신경계에 풍부하게 존재하면서 세포의 안정막 전압을 유지시킨다. 역전사 중합 효소 중합 반응과 면역 세포 화학 염색 방법을 이용하여 자궁내막세포에 존재하는 통로를 조사한 결과, TASK-1, TASK-3, TREK-1, TREK-2 및 TRAAK의 발현이 확인되었다. TASK-3와 TREK-1은 핵을 포함한 세포 전역에 발현
항산화제는 산소의 저장고로서 무혈청 배양액에서 주요한 작용을 하며, 복합배지에서 유용한 첨가제로 알려져 있다. 따라서 본 연구는 한우 체외 수정란의 배양에 있어서 항산화제인 L-cysteine의 작용과 수정란의 발달 단계별 염색체의 분석을 통하여 체외 수정란의 배양 체계를 수립하고자 실시하였다. 한우 난포란의 체외 성숙은 0.1% PVA, 0.1 mM L-cysteine 첨가 시 체외 성숙율은 73.4%, 94.6%으로 각각 나타냈으며, 처리간에 유의적
본 연구에서 돼지 난포란에서 채취된 난모 세포들을 체외성숙 후 형태적으로 선별하거나 극체 방출란을 선별하여 활성화 처리 후 48시간째에 분할란을 선별할 때 배발달율이 어느정도 향상되는지를 검토하였다. 난모 세포를 48시간 성숙 배양 후 형태적 선별과 극체의 방출 유무를 검사하고, 선별된 난모 세포들을 시간 추가 배양한 후 7% ethanol로 활성화시키고 cytochalasin B에 5시간 노출 후 PZM-5 배 양액으로 7일간 배양하였으며, 배양 중