• Journal of Plant Biotechnology
• /
• v.47 no.1
• /
• pp.26-39
• /
• 2020

In maize, immature embryos (IEs) are highly regenerative explants most suitable for producing high frequencies of plantlet regeneration in vitro. Apart from media, explants, and hormones, genotypic variation also influences in vitro characters to a great extent. In the present study, IEs were used to study the distinctive effect of variation of size/stage and hormones in different genotypes on five in vitro characters viz., frequency of callus induction, growth rate of total callus, frequency of E. callus induction, and volume and number of regenerated plantlets. LS medium with different concentrations of 2,4-D (0.5, 1.5, 2.5, 4.0 and 5.0 mg/L) were used to study the former four in vitro characters, and medium with 6-benzylaminopurine and kinetin (0.5 mg/L, each) was used for plantlet regeneration. IEs of 1.0, 1.5, 2.0, 2.5 and 3.0 mm in size were isolated from four inbred lines viz., NM74C, NM81A, NM5883 and NM5884. Two-way ANOVA revealed that explant size and genotypes, as well as hormonal concentrations showed significant effects on in vitro characters. Two millimeter IEs were found to be suitable for in vitro cultures. LS medium with 1.5 mg/L 2,4-D and LS with BAP and Kn (0.5 mg/L, each) were found to be the best hormonal concentrations for callus induction, maintenance, and regeneration, respectively. Among the four genotypes, NM81A and NM5883 yielded more non-embryogenic and Type I E. calli. In contrast, NM74C and NM5884 yielded more highly regenerative Type II calli. Inbred line NM5884 was found to be the best among these four genotypes.

• Animal Bioscience
• /
• v.35 no.2
• /
• pp.177-183
• /
• 2022

Objective: The present study evaluated the efficiency of embryo development and pregnancy of somatic cell nuclear transfer (SCNT) embryos using different source-matured oocytes in Camelus dromedarius. Methods: Camelus dromedarius embryos were produced by SCNT using in vivo- and in vitro- matured oocytes. In vitro embryo developmental capacity of reconstructed embryos was evaluated. To confirm the efficiency of pregnancy and live birth rates, a total of 72 blastocysts using in vitro- matured oocytes transferred into 45 surrogates and 95 blastocysts using in vivo- matured oocytes were transferred into 62 surrogates by transvaginal method. Results: The collected oocytes derived from ovum pick up showed higher maturation potential into metaphase II oocytes than oocytes from the slaughterhouse. The competence of cleavage, and blastocyst were also significantly higher in in vivo- matured oocytes than in vitro- matured oocytes. After embryo transfer, 11 pregnant and 10 live births were confirmed in in vivo- matured oocytes group, and 2 pregnant and 1 live birth were confirmed in in vitro- matured oocytes group. Furthermore, blastocysts produced by in vivo-matured oocytes resulted in significantly higher early pregnancy and live birth rates than in vitro-matured oocytes. Conclusion: In this study, SCNT embryos using in vivo- and in vitro-matured camel oocytes were successfully developed, and pregnancy was established in recipient camels. We also confirmed that in vivo-matured oocytes improved the development of embryos and the pregnancy capacity using the blastocyst embryo transfer method.

• Journal of Embryo Transfer
• /
• v.15 no.1
• /
• pp.33-38
• /
• 2000

This study was undertaken in an effort to select the sire bull in Hanwoo through in vitro fertilization of proven bull semen. It was used for in vitro fertilization that of the 20 proven bull semen with follicular oocytes derived from slaughterhouse ovaries of Hanwoo. The stage of maturation on the time course of bovine cumulus-enclosed oocytes incubated for 24 hours was found the highest(96.4%) than hose of other maturationi time. In vitro fertilization rate of bovine oocytes with proven bull sperm showed from 61.5 to 88.9%. Polyspermy of in vitro fertilized oocytes according to proven bulls were the highest KP 491(61.5%) nothing but KP 486, KP 491 and KP 497.

• Journal of Animal Science and Technology
• /
• v.66 no.3
• /
• pp.577-586
• /
• 2024

The in vitro maturation (IVM) rate of canine oocytes remains low compared to other mammals due to their unique reproductive characteristics. This study aimed to explore the effect of hormone supplementation during the IVM of canine immature oocytes on nuclear maturation and subsequently assess its potential application in canine somatic cell nuclear transfer (SCNT). Immature oocytes were collected and cultured in an IVM medium supplemented with hormones (follicle-stimulating hormone [FSH] and progesterone [P4]) or without hormones (control) for 24 hours. The maturation rates of oocytes in the hormone-treated group (94.92 ± 3.15%) were significantly higher than those in the control group (61.01 ± 4.23%). Both in vitro and in vivo matured oocytes underwent NT to evaluate their utility, and the fusion rates were higher in the in vitro matured group than those in the vivo matured group, not significant between in vivo and in vitro matured group (73.28% and 82.35%, respectively). As a result, 14 fused embryos from the in vitro matured group were transferred into two surrogates, with one surrogate achieving a successful pregnancy and delivering four puppies. Whereas in the in vivo matured group, 85 fused embryos were transferred to 8 surrogate mothers, leading to three surrogates becoming pregnant and delivering one, four, and two puppies. The pregnancy rates were not significant between both groups (50% and 37.50%), but the number of offspring exhibited a significant difference (28.57% and 8.23%). In conclusion, we achieved a remarkable milestone by successfully producing cloned puppies using in vitro matured oocytes, underscoring the feasibility of canine cloning from in vitro recovered oocytes. It is important to note that this study focused only on immature oocytes after ovulation and only during the estrus stage. Further research targeting other stages of the estrous cycle could potentially enhance canine cloning efficiency.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.30 no.1
• /
• pp.129-133
• /
• 2004

The major advantage of the in vitro test is that it is a rapid, objective and cost-effective screening methodology. In vitro tests can provide a formulation tool to identify new fillers that are optimized by combinations of old ones and they can be used to pre-screen protective formulas prior to in vivo testing in humans. Therefore, the accuracy of in vitro SPF measurement is very important. In this study, improvement of application method of samples was tried to improve the accuracy of in vitro SPF measurement. The outer part of Transpore$\^$(R)/ tape was used to apply samples as the substrates and the standard drying time was set at 15 min. The new method, topical applications at light scan areas, results in more accurate and reliable results. This result suggests that more accurate prediction system can be established for in vivo SPF with in vivo SPF measurement.

• Restorative Dentistry and Endodontics
• /
• v.46 no.3
• /
• pp.33.1-33.12
• /
• 2021

Objectives: This study aimed to investigate the efficacy of ionic and non-ionic-based contrast media (in vitro study) and the combinatorial effect of chitosan-based endo-radiopaque solution (CERS) (in vivo study) for visualization of the root canal anatomy. Materials and Methods: In vitro study (120 teeth): The root canal of maxillary premolars and molars (in vitro group 1 and 2 respectively, n = 60 each) were analyzed using 4 different contrast media (subgroups: Omnipaque 350, Iopamidol, Xenetix 350, and Urografin 76; n = 15 each) in combination with 5.25% sodium hypochlorite (NaOCl). Based on the results of the in vitro study, in vivo study (80 teeth) was done to compare Xenetix 350 + 5.25% NaOCl with CERS (in vivo group 1 and 2 respectively, n = 40 each) on maxillary and mandibular premolars and molars. Two endodontists used radiovisiography to assess the depth of ingress and identify the aberrant root anatomy after access cavity preparation, and after initial cleaning and shaping of canals. Kruskal-Wallis test was used for in vitro comparison (p < 0.05), and Wilcoxon signed-rank test and Mann-Whitney U test for in vivo analysis (p < 0.01). Results: In vitro study, Xenetix 350 + 5.25% NaOCl facilitated a significant higher visualization (p < 0.05). For in vivo study, CERS had a statistically significant depth of ingress (p < 0.01), and was efficient in identifying the aberrant root canal anatomy of premolars and molars. Conclusions: CERS facilitates better visualization of the root canal anatomy of human premolars and molars.

• Journal of Embryo Transfer
• /
• v.10 no.3
• /
• pp.219-227
• /
• 1995

This study was investigated to test in vitro-maturation rate of bovine follicular oocytes freezability of in vitro-matured bovine follicular oocytes with different stock solution in Glycerol and Propanediol, freezability of in vitro-rnatured bovine follicular oocytes on cryoprotectants, the viability of in vitro-rnatured bovine follicular oocytes by morphologically normal and FDA staining method. 1. The maturation rates of bovine follicular oocytes classified as grade A, B and C was 88, 63 and 21%, respectively. 2. Freezability of in vitro-matured bovine follicular oocytes on stock solution, TCM-199+5% FCS and m-PBS + 5% FCS was 61%(n=105), 48%(n=62) in $_1$M Glycerol and freeability of in vitro-matured bovine follicular oocytes on stock solution, TCM-199 +5% FCS and m-PBS + 5% FCS was 68%(n=112), 42%(n=57) in 1~2 Propanediol. The results indicate that freezability of in vitro-matured bovine follicular oocytes with different stock solution is important. 3. Freezability of in vitro-matured bovine follicular oocytes on cryoprotectants was Glycerol and PROH was 56%(n=167), 57%(n=169). The results indicate that PROH was superior to Glycerol. 4. The rates of morphologically normal IVM oocytes after thawing of cryopreserved oocytes with Glycerol and PROH were 39%(n=$_1$8), 65%(n=39), respectively. The results indicate that PROH was superior to Glycerol. 5. The fluorescent light intensity after thawing of cryopreserved oocytes classified with Positive, Partial-I, Partial-II, Negative with Glycerol and PROH. The results of FDA-positive 24%, 42%, Partial-I 17%, 10%, Partial- H 20%, 12%, FDA-negative 39%, 37%, and Partial-I, II, respectively.

• Korean Journal of Animal Reproduction
• /
• v.20 no.2
• /
• pp.179-190
• /
• 1996

Sexing and developing from splitted embryos which were fertilized in vitro implicate a possibility of production of the superior and sex controlled individuals. This study was carried out to investigate the production of transferable late blastocysts from in vitro fertilized embryos and to analyze sex by chromosome analysis from same embryos. In results, the ratio of cleavage and fertility of bovine follicular oocytes matured in vitro was 90% in co-cultured with granulosa cells. The competence of embryonic development from in vitro matured and fertilized bovine oocytes was 38% in co-cultured with bovine oviductal epithelial cells. To produce a lot of transferable embryos, therefore, the best conditon of culture system was co-cultured with granulosa cells for immature bovine oocytes and then co-cultured with bovine oviductal eptithelial cells for matured and fertilized oocytes. In chromosome analysis, 93% of in vitro fertilized embryos were very important aspect in chromosome preparation from bovine embryos such as duration of colcemid treatment, weakening of zona pellucida, methods of hypotonic treatment and fixation treatment.

• Korean Journal of Animal Reproduction
• /
• v.20 no.3
• /
• pp.315-322
• /
• 1996

The study was conducted to determine the optimal glucose, superoxide dimutase(SOD) and catalase levels during the in vitro culture of porcine oocytes matured and fertilized in vitro for morulae and blastocyst development. Oocytes were cultured for 0~8 days in TCM-199 medium supplemented with 20% FCS, different glucose, SOD and catalase levels. The results are summairzed as follows ; 1. The in vitro developmental rates of porcine oocytes cultured in TCM-199 medium containing 0.1, 0.3, 0.5, 1.0, 3.0 mM glucose levels 0~3 and 0~8 days after insemination were 22.8, 24.2, 21.9, 20.0, 12.1 and 21.9, 26.7, 25.0, 22.6, 16.7%, respectively. 2. The in vitro developmental rates of porcine oocytes cultured in TCM-199 medium containing 100, 200, 300, 500 $\mu\textrm{g}$/ml SOD levels 0~3 and 0~8 days after insemination were 16.7~23.3 and 16.7~25.0%, respectively. High levels of SOD(500 $\mu\textrm{g}$/ml) significantly reduced the rates of molurae and blastocysts stage(P<0.05). 3. The in vitro developmental rates porcine oocytes cultured in TCM-199 medium containing 100, 200, 300, 500 $\mu\textrm{g}$/ml catalase levels 0~3 and 0~8 days after insemination were 18.8~26.7 and 19.4~28.1%, respectively, and there was significant differences on the development to the molurae and blastocysts stage among the cumulus cells and glucose levels.

• Journal of Embryo Transfer
• /
• v.11 no.3
• /
• pp.225-231
• /
• 1996

This study was conducted to investigate the effects of preincubation time, concentration and exposure time of sperm on in vitro fertilization of porcine follicular oocytes rnatured in in vitro. The results obtained are as follows ; 1. Effect of preincuhation time for porcine sperm capacitation on in vitro fertilization in medium with heparin was investigated. Normal fertilization rate was highest in 15 min(26.4%). However, there were no significant differences among preincuhation times of 5~90 min, 2. Normal fertilization rates of sperm concentrations were 17.0~26.5%, and normal fertilization rate from l$\times$ l05cell /ml concentration was also higher than those of other sperm concentration. 3. Normal fertilization rates of sperm exposure time of 4, 8, 12, 16 and 20 hours were 6.1, 20.8, 27.8, 25.0 and 26.7%, respectively. Normal fertilization rate from sperm exposure time of 12 hours was also higher than that of other sperm exposure times.