• Title/Summary/Keyword: developmental rate

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Effects of various freezing containers for vitrification freezing on mouse oogenesis

  • Kim, Ji Chul;Kim, Jae Myeoung;Seo, Byoung Boo
    • Journal of Animal Science and Technology
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    • v.58 no.3
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    • pp.13.1-13.7
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    • 2016
  • Background: In the present study, various freezing containers were tested for mouse embryos of respective developmental stages; embryos were vitrified and then their survival rate and developmental rate were monitored. Mouse two cell, 8 cell, and blastula stage embryos underwent vitrification freezing-thawing and then their recovery rate, survival rate, development rate, and hatching rate were investigated. Methods: EM-grid, OPS, and cryo-loop were utilized for vitrification freezing-thawing of mouse embryos. Results: It was found that recovery rate and survival rate were higher in the group of cryo-loop compared to those of EM-grid (p < 0.05). Embryonic development rate, two cell embryos to blastocyst, as well as hatching rate were higher in the control group compared to the EM-grid group and OPS group (p < 0.05), yet no difference was noted between the control group and cryo-loop group. Development rate and hatching rate of eight cell morulae and blastocysts were all lower in the treatment groups than the control group whilst hatching rate of blastocysts was higher in the control group compared to the groups of EM-grid and OPS (p < 0.05); although the cryo-loop group was shown to be slightly higher than other groups, it was not statistically significant. Conclusions: In the study, we investigate effects of freezing containers on vitrified embryos of respective developmental stages; it was demonstrated that higher developmental rate was shown in more progressed (or developed) embryos with more blastomeres. There was however, no difference in embryonic development rate was shown amongst containers. Taken together, further additional studies are warranted with regards to 1) manipulation techniques of embryos for various vitrification freezing containers and 2) preventive measures against contamination via liquid nitrogen.

Variation of Transcribed X-linked Genes in Bovine Embryos Cloned with Fibroblasts at Different Age and Cell Cycle

  • Jeon, Byeong-Gyun;Rho, Gyu-Jin
    • Reproductive and Developmental Biology
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    • v.35 no.2
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    • pp.175-183
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    • 2011
  • The present study compared the developmental potential, telomerase activity and transcript levels of X-linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) in the bovine somatic cell nuclear transfer (SCNT) embryos derived from different age and cell cycle of female donor nucleus. In experiment 1, the fusion rate, cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cells was slightly increased in embryos cloned with fetal fibroblasts compared to those with adult fibroblasts, but there was no significantly (p<0.05) differences. Telomerase activity was also similar in blastocysts cloned with fetal and adult fibroblasts. Up-regulated RPS4X and down-regulated MeCP2, XIAP, and XIST transcript level were observed in blastocysts cloned with adult fibroblasts, compared to those with fetal fibroblasts. In experiment 2, the fusion rate, cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cells was significantly (p<0.05) increased in embryos cloned with fetal fibroblasts at early G1 phase of the cell cycle, compared to those of fetal fibroblasts at late G1 phase. DNMT1 transcript was observed to significantly (p<0.05) increased in the fetal fibroblasts at 3 hrs after trypsin treatment of confluent culture. Further, level of telomerase activity and transcribed X-linked genes was also significantly (p<0.05) higher in the early G1 SCNT blastocysts than those of late G1. The results imply that fetal fibroblasts at early G1 phase induces the enhanced developmental potential and up-regulated telomerase activity and X-linked gene, but aberrant transcript pattern of X-linked genes may be displayed in the SCNT embryos.

Effect of Temperature on the Development of the Ash-gray Leaf Bug, Piesma maculata (Insecta, Hemiptera, Piesmatidae) (온도가 두줄명아주노린재의 발육에 미치는 영향 (곤충망, 매미목, 명아주노린재과))

  • Park, Pil Ryoun;Sang Ock Park
    • The Korean Journal of Ecology
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    • v.6 no.1
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    • pp.55-65
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    • 1983
  • The authors reared the ash-gray leaf bug, Piesma maculata in the growth cabinet controlled as temperature groups of $15, 20, 25, 30, 40^{\circ}C$under condition of photoperiod 16L:8D, light intensity $510{\pm}240$ lus, relative humidity $65{\pm}3%$, and analyzed the effects of temperature on the development of the insect. The results are summarized as follow: There are highly significant differences the developmental periods for the temperature groups, and between the developmental periods for the developmental stages. The egg in the temperature of 15 and $40^{\circ}C$ was hatched, but the ecdysis was impossible. The thermal threshold was $12.34^{\circ}C$and the upper lethal temperature $40.39^{\circ}C$. The total developmental periods of egg to adulate in the temperature of 20, 25, 30 and $35^{\circ}C$are 40.52, 22.37, 15.91 and 13.00 days, respectively. That is, the developmental period was decreased, as the temperature was increased. In the developmental period for the developmental stages, the developmental period of egg stage was longer than that of 25, 30 and $35^{\circ}C$, and that of 25。C was longer than that of $35^{\circ}C$. But ther was not significant differences between the developmental periods for the other temperature group. The rate of hatch at$20^{\circ}C$is the greater value as 90%, and the rates of 25, 30 and $35^{\circ}C$ are 79, 79 and 67%, respectively. That is the rate of hatch was decreased, as the temperature was increased. The mortality in the temperature of $35^{\circ}C$ is the greatest value as 68%, and those of 30, 25 and $20^{\circ}C$are 59, 59 and 41%, respectively. That is, the mortality was increased, as the temperature was increased. There was not significantly differences between the developmental period of female and male.

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Studies on the Development of Easy Cryopreservation Technique of Bovine Embryos II. Effects of Equilibration of Cryoprotectants, Temperature and Time of Thawing and 1 Step Straw Method on In Vitro Developmental Rates of Embryos (소 수정란의 간이 동결기법 개발에 관한 연구 II. 내동제의 평형시간, 융해온도, 융해시간 및 1단계 Straw법이 체외발생에 미치는 영향)

  • 김상근;남윤이;현병화
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.103-109
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    • 1997
  • The studies on the carried out to investigate to determine the optimum thawing temperature and equilibration time and 1 step straw method of frozen bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1 $\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as developmental rate on in vitro culture or FDA-test. The results are summarized as followes : 1. The equilibration time on in vitro developmental rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20 min.). 2. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher in vitro developmental rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$. 3. The thawing time on in vitro developmental rates of bovine embryos was attained after short period of time(1~5 min.) in the freezing mediuim higher than long period of time(10min.). 4. The in vitro developmental rates of bovine embryos after rapid frozen-thawing by 1 step straw method in the freezing medium added 1.5M, 2.0M glycerol, DMSO, propanediol and 0.25M, 0.50M, 0.75M, 1.00M sucrose were 12.5~19.4%, 10.0~15.6%, 9.1~13.8% and 6.7~12.9%, respectively.

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The Effect of Cryopreservation Condition on Developmental Rate of Pronuclear Stage Embryos and Vitrification of Mouse Oocytes (생쥐 난자의 유리화 동결과 전핵기 배아의 동결 조건이 배아의 발달에 미치는 영향)

  • Kim, Ji-Chul;Park, Sung-Baek;Nam, Yoon-Sung;Seo, Byoung-Boo;Kim, Jae-Myeoung;Song, Hai-Bum
    • Journal of Embryo Transfer
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    • v.26 no.3
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    • pp.201-207
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    • 2011
  • The present study was performed to investigate the survival and subsequent embryonic developmental rate of immature and mature oocytes after vitrification and pronuclear stage embryos after slow-freezing and vitrification. We have also tried to examine the dependency of concentrations (7.5, 15%) and exposure time (5, 10, 20 min) of ED cryoprotectant on developmental rate of pronuclear stage embryos. The developmental rates of 2-ce1l and blastocyst embryos at mature oocytes were significantly (p<0.05) higher than immature oocytes. After slow freezing, vitrification and thawing of pronuclear stage embryo, the survival and developmental rates of blastocysts and hatched blastocysts were significantly (p<0.05) higher after vitrification than after slow-freezing. On contrary, the developmental rates of 2-cell embryos were significantly (p<0.05) higher after slow freezing than after vitrification. The cryopreservation methods of pronuclear stage embryos vitrified by exposed to 7.5% ED solution for 5 minutes was significantly (p<0.05) higher than other experimental group. The results of our study suggest 1hat the developmental rates of mature oocytes have been more successful than immature oocytes during vitrification. Vitrification was more efficient than slow freezing in case of pronuclear stage embryos. The effective cryopreservation method of pronuclear stage embryos was vitrified by exposed to 7.5% ED solution for 5 minutes.

Study on the Developmental Rate of In Vitro Cultured Cats Oocytes Recovered from Ovaries Collected at Different Stages of the Reproductive Cycle (번식주기의 단계별로 회수한 고양이 난자의 체외수정과 체외발생에 관한 연구)

  • 박상훈;이명헌;김무강;김상근
    • Journal of Embryo Transfer
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    • v.18 no.2
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    • pp.157-161
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    • 2003
  • The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes in vitro maturation of cats oocytes and development of IVM/IVF embryos. The results were summarized as follows : 1. The fertilization and developmental rate of fresh and salts-stored oocytes with and whithout cumulus cells were 65.7%, 17.1% and 28.6%, 8.6% and 57.1%, 13.3%, 23.3%, 3.3%, respectively. The rate of oocytes with cumulus cells(13.3%∼65.7%) was higher than that of denuded oocytes(3.3%∼28.6%). 2. The fertilization and developmental rate of oocytes recovered from ovaries collected at different stages of the reproductive cycle were 68.9%, 44.4%, 48.9% and 17.8%, 8.9%, 12.8%, respectively. 3. The fertilization and developmental rate of oocytes in vitro cultured at different time of incubation(24, 36 and 48 h) were 66.7%, 46.7%, 48.9% and 17.8%, 11.1%, 8.5%, respectively. respectively. The rate of oocytes incubated 24 h(66.7%) was higher than that oocytes incubated 36 and 48 h(46.7%∼48.9%). 4. The fertilization and developmental rate of oocytes treated activation and non-activation oocytes were 57.4%, 31.4% and 22.9%, 11.4%, respectively. The rate of oocytes treated activation was higher than that oocyte treat non-activation.

Developmental Ability of Bovine Embryos Matured and Fetilized In Vitro after Freezing and Thawing to Gastrulation (한우 난포란을 이용하여 생산된 체외수정란의 동결융해후 Gastrulation으로의 체외발생능에 관한 연구)

  • 이명식;장원경;오성종;양보석;박수봉;백광수;정진관;박용윤
    • Korean Journal of Animal Reproduction
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    • v.20 no.1
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    • pp.71-76
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    • 1996
  • This experiment was carried out to investigate the developmental ability of bovine embryos matured and fertilized in vitro to the gastrulation stage. The bovine oocytes were collected from 2∼5mm follicles, matured for 20∼24hrs in 5% CO2 incubator and then fertilized with frozen-thawed semen. On day 9 after IVF and after freezing and thawing the hatching abilities of expanding blastocysts were examined. Cleavage rate and production rate to expanding blastocysts were 59.7%(955/1604) and 20.7%(333/1604), respectively. Hatching rate of day-9 expanding blastocysts was 54%(40/74), that after freezing and thawing was 56%(79/141). Also, developmental ability of hatched blastocysts to the primitive streak stage was 26%(6/23).

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Studies on the Survival Rate of Bisected Porcine Embryos and Immature Oocytes (돼지 분할 수정란 및 미성숙란의 생존율에 관한 연구)

  • 김상근;이명헌;서길웅
    • Korean Journal of Animal Reproduction
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    • v.19 no.2
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    • pp.129-134
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    • 1995
  • This study was carried out to investigate on the survival rate and in vitro developmental rate of bisected porcine embryos and immature oocytes by manipulator and micropipette. Bisected embryos and immature oocytes cultured for 1∼5 days in TCM 199 medium with 20% FCS. Survival rate was defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The survival rate of bisected porcine embryos and oocytes were 26.1%, 22.7%, respectively. The survival rate of bisected embryos and oocytes was significantly lower than that of non-bisection embryos(62.5%). 2. The survival rate of bisected porcine embryos in cultured for 12, 24, 48, 72 hrs with 20% FCS+TCM-199 medium were 26.9%, 19.2%, 19.2% and 11.5%, respectively. 3. The in-vitro developmental rate with and without zona pellucida of bisected porcine embryos by micromanipulator were 30.8%, 25.0%, respectively.

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Effect of temperature on the development of the Common Grass Yellow, Eurema hecabe

  • Kim, Seonghyun;Park, Haechul;Park, Ingyun
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.2
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    • pp.35-39
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    • 2015
  • The developmental responses of insects to temperature are important considerations in gaining a better understanding of their ecology and life histories. Temperature-dependent phenology models permit examination of the effects of temperature on the geographical distributions, population dynamics, and management of insects. Measurements of insect developmental and survival responses to temperature pose practical challenges that depend on the chosen modality, variability among individuals, and high mortality rates near the lower and upper threshold temperatures. Different temperature levels can significantly affect larval development of Eurema hecabe. The development of E. hecabe reared on leaves of Lespedeza cuneata was investigated at three temperature regimes (20, 25, and 30℃), a relative humidity of 60%, and a light:dark photoperiod of 14:10 h. The developmental time from larva to adult was 34.3, 20.6, and 17.9 d at temperatures of 20, 25, and 30℃, respectively. Pupal rate was 47.6%, 47.6%, and 61.9% at temperatures of 20, 25, and 30℃, respectively. The developmental threshold temperature estimated from larva to pupae was 8.1℃ with 381.7 degree-days. There is an increasing need for a standardized manual for rearing this butterfly species based on adequate knowledge of its ecology.

Studies on the Survival and In Vitro Developmental Rates after Bisection of Bovine Embryos (소 초기배 분할후 생존성과 체외발생율에 관한 연구)

  • ;Y. Noriko
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.275-280
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    • 1997
  • This study was carried out to investigate on the survival and in vitro developmental rates of bisected bovine embryos by microblade, micropipette and pronase methods. Bisected embryos cultured for 1∼7 days in TCM-199 media with 10 FCS+hormones. Survival and in vitro developmental rates was defined on in vitro culture or FDA-test. The results are summarized as follows ; 1. The survival and in vitro developmental rates of bisected bovine embryos by microblade, micropipette and pronase methods were 22.2, 16.7, 15.0% and 22.2, 23.3, 18.8%, respectively. In vitro developmental rate of bisected bovine embryos was significantly lower than that of non-bisection embryos(27.8% and 25.0%). 2. In vitro developmental rates of bovine embryos bisected for 1, 2, 4, 8, 16 cells stages during in vitro culture in 10% FCS+TCM-199 media were 25.0, 20.0, 20.0, 15.0 and 6.7%, respectively. 3. In vitro developmental rates of intact and free-zona pellucida of bisected demi-embryos during in vitro culture in 10% FCS+TCM-199 media were 25.6, 16.7%, respectively. 4. In vitro developmental rates of biopsied embryos and biopsied blastomeres during in vitro culture in 10% FCS+TCM-199 media were 20.0, 11.1%, respectively.

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