• 제목/요약/키워드: Embryo production

검색결과 693건 처리시간 0.03초

Detection of the SRY Transcript and Protein in Bovine Ejaculated Spermatozoa

  • Li, Chunjin;Sun, Yongfeng;Yi, Kangle;Li, Chengjiao;Zhu, Xiaoling;Chen, Lu;Zhou, Xu
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제24권10호
    • /
    • pp.1358-1364
    • /
    • 2011
  • The sex-determining region on the Y (SRY) gene is important in mammalian sex determination and differentiation. We report a study of the abundance of SRY gene products in bovine ejaculate. RT-PCR experiments using RNA extracted from bovine spermatozoa with SRY-specific primers yielded a 456 bp product, but the amount of SRY mRNA in sperm was lower than that in the testes (p<0.01). A protein of approximately 27 KDa was detected by western blotting. The SRY transcript was detected in the midpiece of approximately half the spermatozoa by in situ hybridization, and the SRY protein was detected in the heads of half the spermatozoa by immunofluorescence, indicating that SRY mRNA and protein may only be present in Y-bearing spermatozoa. These results suggest that the SRY transcript and protein are present in bovine ejaculated Y-sperm. The roles of the SRY gene in spermatogenesis, sperm motility, and the sperm-oocyte interaction merit further investigation.

Effect of Alpha-Linolenic Acid on Oocyte Maturation and Embryo Development in Pigs

  • Lee, Ji-Eun;Hwangbo, Yong;Kim, Hwa-Young;Lee, Won-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • 한국발생생물학회지:발생과생식
    • /
    • 제21권2호
    • /
    • pp.205-213
    • /
    • 2017
  • The aim of this study was to determine the effect of additional alpha-linolenic acid (ALA) supplementation during in vitro maturation (IVM) and culture (IVC) on nucleic maturation and embryo development of pigs. Cumulus-oocyte complexes (COCs) were incubated in IVM medium containing different concentration of ALA (25, 50 and $100{\mu}M$) for 44 h. After in vitro maturation, nuclear maturation of oocytes were evaluated by aceto-orcein stain. Mature oocytes with $50{\mu}M$ ALA were fertilized and cultured in IVC medium with ALA (25, 50 and $100{\mu}M$) during early-embryogenesis (48 hours after fertilization). Then, embryos were cultured with $25{\mu}M$ ALA during early embryogenesis and/or late embryogenesis (120 hours after early-embryogenesis). In results, oocyte maturation were significantly increased by $50{\mu}M$ ALA treatment groups compared with control groups (p<0.05). Treatment of $25{\mu}M$ ALA during early-embryogenesis enhanced cleavage rate of embryo compared with other groups (p<0.05), whereas formation and total cell number of blastocyst had no significant difference. Similarly, cleavage rate of embryos were increased by $25{\mu}M$ ALA supplement during early- or late-embryogenesis than ALA treatment both stage of embryogenesis (p<0.05), but did not influence to blastocyst formation. Interestingly, total cell number of blastocyst were enhanced in ALA treatment group during early-embryogenesis. These findings indicated that ALA supplement enhance the nuclear maturation of oocyte and embryo development, however, excessive ALA could negatively influence. Therefore, we suggest that ALA is used for improvement of in vitro production of mammalian embryo and further study regarding with functional mechanism of ALA is needed.

Embryo Survival on Day 25 of Generation in the Gilt is Not Affected by Exogenous Progesterone but is Correlated with Levels of Insulin-Like Growth Factor-I (IGF-I) mRNA in the Uterus

  • Yu, Z.;Gordon, J.R.;Kirkwood, R.N.;Thacker, P.A.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제12권6호
    • /
    • pp.862-867
    • /
    • 1999
  • The present study was undertaken to determine the effect of administration of exogenous progesterone early in gestation on uterine levels of IGF-I mRNA and on embryo survival at day 25 of gestation in the pig. Forty-one prepubertal gilts were induced into oestrus with PG600 and artificially inseminated at their subsequent naturally occurring oestrus. Gilts were then randomly assigned to one of three groups. Gilts in the two treatment groups were injected intramuscularly with 50 mg of progesterone either from day 2 to 14 (N=14) or from day 4 to 14 (N=15) after breeding while those in the control group (N=12) were given corn oil (0.5 ml) from day 2 to 14. Between days 25 and 28 of gestation, gilts were slaughtered and reproductive tracts were recovered. Endometrial tissue (1 g) was collected and analysed for IGF-I mRNA levels using a reverse transcription-polymerase chain reaction, Progesterone treatment, starting either on day 2 or 4 after breeding, neither significantly increased embryo survival rate by day 25 of gestation nor altered IGF-I mRNA levels in uterine tissue. However, across all samples, the IGF-I mRNA level in the uterus was highly correlated with embryo survival rate (r=0.8193, p<0.01), supporting the involvement of IGF-I in the regulation of porcine embryo development.

Cryopreservation of Embryo by Concentration of Ethylene Glycol and Day 6, 7, 8, 9 Embryo in Korean Native Cattle (Hanwoo)

  • Park, Joung-Jun;Yoo, Han-Jun;Choi, Hye-Won;Cheong, Ki-Soo;Kim, Ji-Tae;Park, Choon-Keun;Yang, Boh-Suk
    • 한국수정란이식학회지
    • /
    • 제27권4호
    • /
    • pp.253-258
    • /
    • 2012
  • This study was carried out to effects of ethylene glycol concentration, sucrose and culture day of in vitro production embryo on slow-down freezing in Hanwoo. 6, 7, 8 and 9 day embryos produced in vitro were frozen using 1.8M EG+0.1M sucrose, 1.8M EG+0.5% BSA and 1.5M EG+0.1M sucrose media. Survivability was confirmed after frozen-thawed 24 and 48h and ICM, TE cell number were counted by Hoechst 33342 and PI staining after frozen-thawed 24h. As a result, 1.8M EG+0.1M sucrose group was most significantly (p<0.05) higher compared with the other treatment groups on survivability, TE and total cell number after frozen-thawed 24h ($94.2{\pm}2.6%$, $94.67{\pm}3.4$ and $129.67{\pm}5.5$). ICM number did not found significant (p<0.05) differences between the three treatment groups. in 6, 7, 8 and 9 day of embryos using three types of freezing media, frozen-thawed, 1.8M EG+0.1M sucrose groups with embryos cultured 8 day was significantly (p<0.05) highest survivability to $98.3{\pm}1.7%$ after frozen-thawed 24h. 1.5M EG+0.1 sucrose group with embryos cultured 9 day was significantly higher survivability than group of embryos cultured 8 day after frozen-thawed 24 and 48h. In conclusion, 1.8M EG+0.1M sucrose media is considered to be effective to cryopreservation of embryos cultured 8 and 9 day.

한우 수정란의 동결보존 및 쌍자생산에 관한 연구 I. 동결 수정란의 이식과 자우 생산 (Studies on Embryo Cryopreservation and Twinning by Embryo Transfer of Korean Native Cattle I. Transfer of Frozen-thawed Embryos and Production of Calves)

  • 손동수;김일화;이호준;서국현;이동원;류일선;이광선;전기준;손삼규
    • 한국수정란이식학회지
    • /
    • 제12권1호
    • /
    • pp.75-90
    • /
    • 1997
  • This study was carried out to establish the techniques for producing the calves of genetically superior Korean Native cattle by transfer of frozen-thawed embryos. The effects of some factors related to embryo recovery following superovulation and pregnancy rate following transfer of frozen-thawed embryos were evaluated. Also calving state was investigated. The results obtained were as follows ; The mean number of total and transferrable embryos recovered per superovulated cow was 8.72 and 4.90, respectively, from a total of 72 superovulations using 34 donor cows. There were no significant differences in the number of total or transferrable embryos recovered per superovulated cow between products of follicle stimulating hormone (FSH), years, seasons, and collection numbers. The pregnancy rate was found 44.44% following transfer of frozen-thawed embryos of Korean Native cattle to a total of 180 recipient cows including 82 Angus, 27 Charolais, 62 Hereford and 9 Korean Native cows. The pregnancy rate was significantly (P<0.05) higher in the transfer of excellent (42.99) and good embryos (40.17%), compared with fair (5.90%) grade embryos. And the pregnancy rate was significantly (P<0.05) higher in the transfer of embryos of morula stage (43.86%) than blastocyst stage (15.51%). But there were no significant differences in pregnancy rates between natural and induced estrus estrus asynchrony of 1 days, breeds, and parities of recipient cows. The normal calving rate of 80 pregnant cows following transfer of frozen4hawed em-bryos was 87.5% and the other 10 pregnant cows showed abortion during the period from pregnancy diagnosis at 50~60 days to calving. The average gestation length of normally delivered recipients was 288.50 days and the average birth weight of 70 calves born was 24.22 kg. The gestation length was significantly (P<0.05) shorter in the recipients delivering female calves (286.70 days) than males (289.39 days). But there were no significant differences in gestation tength and birth weight of calves born between the recipient breeds.

  • PDF

한우의 수정란이식 효율성 향상을 위한 생체난포란 채취에 관한 연구 (Study on Ovum-pick up for Improvement of Embryo Transfer Efficiency in Hanwoo Cows)

  • 조상래;강성식;김의형;이석동;이명숙;양병철
    • 한국수정란이식학회지
    • /
    • 제32권3호
    • /
    • pp.147-151
    • /
    • 2017
  • Commercial applications of OPU/IVP were to produce embryos and calves from high genetic cows. The aim of this present study was to compare the number of recovered oocytes and cultured In vitro produced embryos from Ovum Pick-up (OPU). OPU derived embryo production was carried out of oocytes by ultrasonographic guided follicular aspiration and then produced in vitro produced blastocysts by IVP culture system. In result, the rate of recovered oocytes was obtained 612 (57.2%) and 451(73.7) G1+G2 grade oocytes. No difference of recovered rate (51.1~62.1%) was seen in six donor. The rate of cleavage and blastocyst development were obtained 320 (70.9%) and 78 (24.4%) that was $3.3{\pm}0.4$ cleaved embryo and $0.9{\pm}0.2$ blastocysts per session. Cleavage rate of OPU oocytes in No. 6 donor was 90.6%, significantly (P<0.05) higher than that in the other donors, However, blastocysts was similar (25.8~30.0%). In conclusion, limited numbers of OPU oocytes had competent development when cultured in SOF culture medium.

한우 과배란처리시 CIDR 처리와 미처리가 수정란 생산에 미치는 영향 (The Effect of Disregarding of Heat Detection on Embryo Production in Superovulation of Hanwoo)

  • 최선호;손동수;류일선;조상래;한만희;김현종;최창용;김영근
    • 한국수정란이식학회지
    • /
    • 제20권3호
    • /
    • pp.311-315
    • /
    • 2005
  • 본 연구는 한우를 공란우로 이용시 효율을 최대화하고, 과도한 호르몬의 투여를 억제하며, 그로 인하여 공란우의 경제수명을 연장하고자 과배란 처리시 차기 발정을 고려하지 않고 과배란을 유도하였을때 공란우의 반응과 수정란의 생산율을 조사하였으며, 그 결과는 다음과 같다. 1. 한우 경산우 및 미경산우를 자연발정을 확인하고 차기발정 5일전에 FSH를 1일 2회 4일 간 투여한 것과 발정을 확인하지 않고 CIDR를 이용하여 동일한 호르몬을 방법으로 과배란을 처리하였을 때, 반응율은 각각 $82.2\%,\;89.7\%$였으며, 유의적인 차이는 인정되지 않았다. 2. 한우 경산우 및 미경산우를 자연발정을 확인하고 차기 발정 5일전에 FSH를 1일 2회 4일간 투여한 것과 발정을 확인하지 않고 CIDR를 이용하여 동일한 호르몬을 같은 방법으로 과배란을 처리하였을 때, 수정란의 회수율은 각각 7.7개, 10.5개를 회수하였고, 그 중 이식가능 수정란은 각각 3.4개, 3.8개를 나타냈으며, 두 처리간에 유의적인 차이를 나타냈다(p<0.05). 이상의 결과로 한우 공란우는 경산우 및 미경산우를 발정주기를 확인하지 않고도 과배란처리시 반응을 확인할 수 있었으며, 수정란의 회수에 있어서도 적절하게 채취할 수 있으므로 한우를 공란우로서 과도한 호르몬으로부터 보호할 수 있으며, 그에 따라 경제수명을 연장할 수 있을 것으로 사료된다. 또한 고능력 한우를 이용한 수정란 이식의 활성화에 유용한 방법이며, 농가에도 확산하여 소득향상에 기여할 수 있을 것으로 기대된다.

Pregnancy Rate of In Vitro Produced Korean Cattle Embryos according to Transport Time Course

  • Park, Hyo-Young;Kim, Eun-Young;Kim, Young-Hun;Mun, Seong-Ho;Oh, Chang-Eon;Han, Young-Joon;Kim, Nam-Hyung;Lee, Sung-Soo;Ko, Moon-Suck;Riu, Key Zung;Park, Se-Pill
    • Reproductive and Developmental Biology
    • /
    • 제33권4호
    • /
    • pp.257-262
    • /
    • 2009
  • This study was to investigate pregnancy rate of IVM/IVF/IVC Korean cattle (registered in government) embryos according to transport time course. For the production of embryos, oocytes recovered from slaughtered excellent grade cow and highly motile frozen-thawed bull semen (purchased from LIMC, KPN#497) was used. In vitro produced embryos were cultured in CR1aa medium for 8 days and some of them were frozen. The rate of average cleavage (>2-cell) was 83.0% (308/371) and blastocyst rate at day 8 was 34.7% (107/308). Among in vitro produced blastocyst embryos at day 8, most healthy embryos were freshly transferred on production day and some frozen embryos were direct transferred on appropriate day. These embryos were produced in a laboratory, embryo transfer (ET) was planned in 10 areas of the remote island (Jeju) from the laboratory by airplane. Thus, we examined the pregnancy rate in recipient cow according to embryo of transport time course before ET. From embryo transferred 44 recipient cows, overall pregnancy was 40.9% (18/44), these 18 cows were all calved [single, 94% (17/18); twin, 6% (1/18)] and total embryo implantation rate was 26% (19/66). Comparing transport time in the base of 6 hr, pregnancy rate in ET group required less 4 hr (60%, 9/15) was significantly higher than that required more 6 hr (26.3%, 5/19). In direct ET of freezing embryos, the pregnancy rate was 40% (4/10). However, it was difficult to find the meaning of temperature, pH and corpus luteum quality of recipients on comparison of pregnancy rate. When the cell death level of embryos according to storage time in thermos (straw container) before ET was measured by TUNEL staining, apoptotic index was increased with storage time-dependent. These results demonstrated that long distance transfer of IVM/IVF/IVC embryos is possible and the time of embryo transport is very important for the pregnancy rate on field trial.

한우 육질등급이 난포란의 배반포 체외생산에 미치는 영향 (Correlation of Hanwoo (Korean Native Cattle) Carcass Classification and Oocyte Donor for Blastocyst Production In Vitro)

  • 김강식;이홍철;박용수;김소섭;박흠대
    • 한국수정란이식학회지
    • /
    • 제30권3호
    • /
    • pp.161-170
    • /
    • 2015
  • These studies were conducted to establish the practical Hanwoo (Korean native cattle) improvement system through the combining of embryo transfer technology and confirming individual Hanwoo oocyte culture system and to investigate that correlation of Hanwoo carcass classification (intramuscular marbling) and oocyte donor for blastocyst production in vitro. In case of Hanwoo, the carcass meat quality grades were divided to grade $1^{{+}{+}}$, $1^{+}$, 1, 2, and 3 depends on fat distribution of longest muscle cross-sectional surface. As results, the numbers of follicular oocytes collected from individual fundamentally-registered Hanwoo yielded $1^{{+}{+}}$, $1^{+}$, 1, 2 and 3 meat quality were 9.5, 9.4, 8.5, 8.8 and 8.8 per ovary, respectively. The numbers of retrieval oocyte from follicles were significantly higher in the cattle yield $1^{{+}{+}}$, $1^{+}$ meat quality than in the cattle yield 1, 2 and 3 meat quality (p<0.05). The rates of blastocyst formation were 18.2, 21.3, 29.4, 30.9, and 31.5% in the cattle yield $1^{{+}{+}}$, $1^{+}$, 1, 2 and 3 meat quality of after in vitro maturation, respectively. It was significantly lower in the cattle yield $^{{+}{+}}$ and $1^{+}$ meat quality than in the cattle yield 1, 2 and 3 meat quality (p<0.05). In order to evaluate embryos quality, TUNNEL assay was conducted for each meat quality grade using blastocyst stage embryo on day 8. The results showed that apoptosis cell number was higher tendency in the cattle yield $1^{{+}{+}}$and $1^{+}$ meat quality (81 and 79, respectively) than in the cattle yield 1, 2 and 3 meat quality (51, 48 and 50, respectively) but there was no statistical significance in each group. After embryo transfer, the conception rate of recipient was 53.5 (23 out of 43), 52.1 (38 out of 73) and 58.0% (58 out of 100) in the meat quality of 1, $1^{+}$ and $1^{{+}{+}}$, respectively. These results showed that the conception rate was significantly higher in the cattle yield 1 meat quality than in the cattle yield $1^{{+}{+}}$, $1^{+}$, 2, and 3 meat quality (p<0.05). In summary, these results indicate that the application of confirming Hanwoo individual oocyte culture system and embryo transfer technology can make good use of the genetic resources conservation and improvement of Hanwoo. Relevance of the meat quality grade and reproductive ability of carcasses of Hanwoo will be considered to be one of the effective means for the associated research with obesity and reproduction.

Dysfunctional pancreatic cells differentiated from induced pluripotent stem cells with mitochondrial DNA mutations

  • So, Seongjun;Lee, Song;Lee, Yeonmi;Han, Jongsuk;Kang, Soonsuk;Choi, Jiwan;Kim, Bitnara;Kim, Deokhoon;Yoo, Hyun-Ju;Shim, In-Kyong;Oh, Ju-Yun;Lee, Yu-Na;Kim, Song-Cheol;Kang, Eunju
    • BMB Reports
    • /
    • 제55권9호
    • /
    • pp.453-458
    • /
    • 2022
  • Diabetes mellitus (DM) is a serious disease in which blood sugar levels rise abnormally because of failed insulin production or decreased insulin sensitivity. Although many studies are being conducted for the treatment or early diagnosis of DM, it is not fully understood how mitochondrial genome (mtDNA) abnormalities appear in patients with DM. Here, we induced iPSCs from fibroblasts, PBMCs, or pancreatic cells of three patients with type 2 DM (T2D) and three patients with non-diabetes counterpart. The mtDNA mutations were detected randomly without any tendency among tissues or patients. In T2D patients, 62% (21/34) of iPSC clones harbored multiple mtDNA mutations, of which 37% were homoplasmy at the 100% mutation level compared to only 8% in non-diabetes. We next selected iPSC clones that were a wild type or carried mutations and differentiated into pancreatic cells. Oxygen consumption rates were significantly lower in cells carrying mutant mtDNA. Additionally, the mutant cells exhibited decreased production of insulin and reduced secretion of insulin in response to glucose. Overall, the results suggest that screening mtDNA mutations in iPSCs from patients with T2D is an essential step before pancreatic cell differentiation for disease modeling or autologous cell therapy.