• Current Research on Agriculture and Life Sciences
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• 제8권
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• pp.51-58
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• 1990

본(本) 연구는 micromanipultor를 이용하여 생쥐의 8세포기배와 상실배 그리고 배반포기배를 양분(兩分)후 생존성을 검토하고, 또한 배반포기배를 양분후, 선별(選別)및 배양 과정없이 암컷 생쥐에 이식(移植)하는 경우 새끼쥐 생산의 가능성을 검토하고자 수행된 것이다. 그 경과를 요약해 보면 다음과 같다. 1. 생쥐의 8세포기배(細胞期胚)와 상실배(桑實胚)를 양분(兩分)하여 M2에 배양한결과 각각 64%, 81%가 배반포기배(胚盤胞期胚)까지 발생(發生)하였다. 2. M2 배양액에서 발생시킨 배반포기배를 Ham's F-10에서 배양한 결과 8세포기배에서는 86%, 상실배에서는 90%가 정상적으로 outgrowth 되었다. 3. 배반포기배를 배양 과정없이 바로 양분(兩分)하여 Ham's F-10에서 배양한 결과 97%가 정상적으로 outgrowth 되었으나 암컷생쥐에 이식한 결과 산차는 얻지 못하였다.

• 한국가축번식학회지
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• 제17권1호
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• pp.57-68
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• 1993

This stduy was carried out to find a reliable method for the production of in vitro fertilized embryos having more excellent development capacity and freezability in the rabbit and cattle. The greatest number of rabbit oocytes was recovered 6hrs after HCG injection(P<0.05). The maturation rate in vitro was slightly higher in the oocytes(6-h-oocytes) from 6h than those (8-h-oocytes)from 8 hrs after HCG injection and the beneficial effect of FSH during oocyte maturation was significantly great in the oocytes from large follicles. The cleavage rate into 2-to-6-cell stage was not differ between the 6-h-oocytes and 8h-oocytes, but the cleavage of these oocytes was greatly promoted by FSH addition to maturation medium and the cleavge of 8-h-oocytes matured without FSH was significantly low. The embryo development into 16-cell to morula was not promoted by the co-culture with rabbit oviduct epithelial cells. The freezability by embryo stages was ovidusly high at 4-cell and morula stage in 6-h-oocytes and the viability of 16-cell embryos from 8h-oocytes was similar to that of morula stage. The implantation sites after surgical tranfer of fresh rabbit embryos were not implanted. In bovine experiment, the in vitro development into 16-cell and morula after in vitro maturation and fertilization in the follicular oocytes was slightly improved by the co-culture with granulosa cells compared to that with oviduct epithelial cells and the frozen-thawed viability rate of these embryos ranged from 14 to 40%. The excellent fresh embryos were transferred nonsurgically to 6 recipients, but were not pregnant.

• 한국수정란이식학회지
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• 제4권1호
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• pp.28-34
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• 1989

These experiments were carried out to determine the effect of cell stage in embryo bisection on the sub-Sequent in vitro and in vivo development in mouse. The embryos of ICR mouse were microsurgicaily bisected at 2-cell, 4-cell, 8-cell, morula and blastocyst stage using a microsurgical blade attached a micromanipulator. These demi-embryos without zona pellucida were cultured up to blastocyst stage and transferred to pseudopregnant mice, and the development of these demi-embryos was compared with the results of intact embryos of the corresponding cell stage. The successful rate of mouse embryo bisection at 4-cell stage (59.0%) was significantly (p <0.05) lower than those at 8-cell (75.6%), 2ce11 (80.7%) or morula stage (84.8%), and highest at blastocyst stage (95.7%). When the bisected embryos without any damage from microsurgery were cultured in vitro up to blastocyst,the in vitro de'velopment of demi-embroys bisected at morula to blastocyst was 91.6 to 95.3%, which was similar to the culture result of intact embryos of corresponding stage. However, the in vitro development of demi-em-bryos bisected at 2- to 8-cell stage was signiflcantiy (p <0.05) lower.The post-transfer implantation rate of demi-embryos developed in vitro to eu-blastocyst were 19.6 and 25.4% in demi-embryos bisected at morula and blastocyst stage,respectively and not significantly (P <0.05)different from the result of intact embryos of the same stage. However, the implantation rates of demi-embryos bisected at 2- or 8-cell stage were significantly (P <0.05) lower than the result from the intact embryos of the corresponding stage.

• 한국가축번식학회지
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• 제20권3호
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• pp.233-238
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• 1996

This study was conducted to investigate the effects of cryoprotective agents and thawing temperature on the survival rate of the bisected embryos of mouse. The results of this study were summairzed as follows: 1. In in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed, the rates of normally developed bisected morula which was denuded were 31.7, 39.1, 28.0 and 23.1%, respectively. 2. The survival rates of bisected morula encased into the zona pellucida in in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed were 72.4, 68.7, 64.0 and 59.5%, respectively. 3. The survival rates of bisected denuded blastura in in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed were 48.3, 44.8, 32.1 and 28.6%, respectively. 4. The survival rates of bisected blasturae encased into the zona pellucida in in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed were 73.6, 67.4, 53.0 and 49.1%, respectively. 5. In in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed at room temperature, the rates of the normally developed bisected morula which was encased into the zona pellucida were 67.1, 62.3, 57.7 and 53.0%, respectively, and those of the bisected blasturae encased into the zona pellucida were 70.8, 65.4, 56.6 and 52.1%, respectively. 6. The survival rates of bisected morula which was encased into the zona pellucida in in vitro culture after forzen with DMSO, glycerol, ethylene glycerol or propanediol and thawed at 37$^{\circ}C$ were 74.3, 71.3, 63.9 and 57.4%, respectively, and those of bisected blastura encased into zona pellucida were 76.0, 69.1, 61.1 and 56.1%, respectively.

• 한국가축번식학회지
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• 제14권1호
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• pp.43-49
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• 1990

To improve the freezing techniques of animal embryos using vitrification solution as a cryoprotectant rabbit embryos, by cell stages, dehydration temperature and dehydration temperature and dehydratin time, were frozen-thawed and cultured. Following are the main results obtained. 1. The damage rate of zona pellucida after thawing was higher(13.6%) when the cell stage of embryos was less than 4 cells than when the cell stage was 8~16 cell or morula. The damage rate was higher when the dehydration temperature was 4$^{\circ}C$ than -3$0^{\circ}C$ or -50~-8$0^{\circ}C$. The zona pellucida was damaged more when dehydrated for 5 min than when dehydrated for 10~15 min. 2. After being cultured for 72 hours, 5.3% of 4 cell(or less) embryos were developed to morula, while 86.4% of morula embryos were developed further. 3. More percentage of embryos(73.2%) was developed when dehydrated at -3$0^{\circ}C$ than when dehydrated at 4$^{\circ}C$ at -5$0^{\circ}C$~-8$0^{\circ}C$. 4. The hatching rate was higher when dehydrated for 5 min. When the embryos were dehydrated for 10~15 min and cultured for 24 hours, they were not even developed or development was not good in later stages.

• 한국가축번식학회지
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• 제11권3호
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• pp.218-222
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• 1987

This study was conducted using inbred ICR mice to investigate the sex-ratio of preimplantation mouse embryos. For the investigation of sex-ratio of mouse embryos, the karyotype of embryos collected at 70-72, 74-76, 78-80 and 82-84 hr after HCG injection was analyzed by chromosomal analysis. Eight-cell embryos were cultrued up to blastocyst stage, then divided them into three groups(fast-, intermediate- and slow-) according to the blastocoel formation. The sex-ratio was also investigated by chromosomal analysis. 1. The highest apperance of eight-cell and morula was observed at the embryos collected respectively at 66-68 hr(84.6%) and 82-84 hr(79.3%) compared to any other group. 2. The successful rate of embryos sexing at 4-, 8-cell and morula stage were 23.1% (3/13), 42.1%(138/328) and 32.6%(47/141), respectively. The respective sex ratios (female vs male) of 4-, 8-cell and morula were 66.7:33.3, 49.3:50.7 and 39.5:60.5. 3. Of the 476 eight-cell embryos cultured in vitro, 427(89.7%) embryos were developed to the blastocysts and the number of fast-, intermediate- and show-developing embryos were 139, 144 and 144, respectively. 4. Female to male ratios fo fast-, intermediate- and slow-developing group were 23.0:77.0, 55.2:44.8 and 73.8:26.2, respectively. Significantly higher (P<0.05) number of female (48/65;73.8%) was observed in the group of slow-developing embryo than that out of total number of embryos(82/188;43.6%).

• 한국수산과학회지
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• 제45권5호
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• pp.480-485
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• 2012

The hemocytes of the tunicate Halocynthia roretzi are classified into six types based on their size, cellular form, and fine structure of the cytoplasmic granules: hyalinocytes, granulocytes, phagocytes, nephrocytes, morula cells, and multi-vacuole cells. Based on cell size, they are ordered multi-vacuole cells ($7.5{\mu}m$)>nephrocytes ($7.1{\mu}m$)>phagocytes ($6.8{\mu}m$)>granulocytes ($6.1{\mu}m$)>morula cells ($5.7{\mu}m$)>hyalinocytes ($5.4{\mu}m$). The proportion of hemocytes is ranked in the order multi-vacuole cells (54.8%)>nephrocytes (16.9%)>granulocytes (9.9%)>morula cells (8.8%)>phagocytes (6.1%)>hyalinocytes (3.5%).

• 한국가축번식학회지
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• 제16권3호
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• pp.217-224
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• 1992

These studies were carried out ot investigate on the transferred embryo development following ultrarapid frozen for 8-cell and morula of in vitro fertilization mouse embryos. The post-thaw embryo survival was evaluated and compared by cell stage of embryos and by equilibration time before ultrarapid freezing. The results obatined were summerized as follows: 1. The effects of equilibration time of 3 vs. 6 minutes before ultrarapid freezing and after thawing on the morphological survival and the viability of 8-cell and morulas embryos were not significant. 2. When the ultrarapid frozen-thawed 32 eight-cell and 33 morula embryos, and 30 fresh blastocysts were transferred to pseudopregnant recipient mice, the number of normal offsprings produced were 9(28.1%), 14(42.4%) and 18(60.0%), respectively. From the above resutls, it was concluded that the optimal conditions of pH osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOsm and 120 min., respectively,a nd somewhat high conception rate might be resulted from transfer of frozen embryos of morula stage and fresh embryos of blastocyst stage.

• 한국임상수의학회지
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• 제19권1호
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• pp.73-79
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• 2002

To investigate the usability of frozen canine embryos for embryo transfer in the dog, 19 donors, 3 recipients, and 6 male dogs were used for the experiment. Natural mating or artificial insemination was performed for breeding the bitches in natural estrus. Vaginal smear test along with progesterone titre test were performed to detect the appropriate mating time and the bitches were bred twice during 3-6days following LH surge. Embryo collection was done on 8, 9-11, 12-13 days after the second mating to collect morula and blastocyst. Embryos were frozen using a programmable freezer and preseued in LNE tank. Embryos were thawed in 37$^{\circ}C$ water for 15 seconds and transferred into each uterine horn within 30 minutes. Embryos were collected from 13 bitches of 19 donors(68.4%) and the collected embryos were from between 9 and 13 days after 2nd mating. Embryos were produced both by natural mating(60.0%, 9115) and AI with frozen semen(100.0%, 4/4). Embryos were collected from the donors weighed between 2.5 and 30 kg and their age was from 1.5 to 3 years. 52 embryos were collected from 13 donors and the mean number of embryos was four. The stage of embryos was from 2-cell to gastrula and morulae were colledted mostly from 10 to 11 days after 2nd mating. Embryos were collected evenly from each uterine horn and the rate of embryo collection for the number of corpus luteum was 83.9%. Embryos were transferred to 3 recipients(morula 8, blastocyst 1, gastrula 8), however, no offspring was produced.

• 한국동물생명공학회지
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• 제36권4호
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• pp.307-313
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• 2021

Traf4 (Tumor necrosis factor Receptor Associated Factor 4) is a member of the tumor necrosis factor receptor (TNFR) - associated factors (TRAFs) family. TRAF4 is overexpressed in tumor cells such as breast cancer and associated with cytoskeleton and membrane fraction. Interestingly, TRAF4 was localized with tight junctions (TJs) proteins including OCLN and TJP1 in mammary epithelial cells. However, the expression patterns and biological function of Traf4 were not examined in preimplantation mouse embryos although Traf4-deficient mouse showed embryonic lethality or various dramatic malformation. In this study, we examined the temporal and spatial expression patterns of mouse Traf4 during preimplantation development by qRT-PCR and immunostaining, and its biological function by using siRNA injection. We found upregulation of Traf4 from the 8-cell stage onwards and apical region of cell - cell contact sites at morula and blastocyst embryos. Moreover, Traf4 knockdown led to defective TJs without alteration of genes associated with TJ assembly but elevated p21 expression at the KD morula. Taken together, Traf4 is required for TJs assembly and cell proliferation during morula to blastocyst transition.