• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.97-97
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• 2002

Achieving of in vitro development for mammalian premature spermatogenic cells are very difficult. In-vitro culture of spermatogenic cells were then initiated in an effort to try to study in vivo spermatogenesis and to understand its molecular events. Recently, the morphogenetic changes of spermatocytes or spermatid by in-vitro culture system were achieved. (omitted)

• Mycobiology
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• v.34 no.1
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• pp.1-6
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• 2006

A Cordyceps species was found with a Beauveria anamorph state on larval insect cadavers on Obong mountains in Gangwon Provinces, Republic of Korea. Cultures from discharged ascospores formed an anamorph identifiable as Beauveria bassiana. This teleomorph-anamorph connection was also confirmed by the in vitro production of fertile ascomata from conidial cultures with morphology like that of field-collected specimen. This is the first report of in vitro production of a teleomorph for any Beauveria species. The Cordyceps species has been conspecified as Cordyceps bassiana, a species described from China with B. bassiana anamorph.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.919-927
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• 1996

In the last few years, methods for in vitro culture of early embryo stages from oocytes matured and fertilized in vitro using suitable cell culture systems have been established. But the factors affecting pregnancy rates following transfer of bovine embryos produced in vitro were not evaluated enough. So this study was performed to investigate the effects of quality and stage of embryos, parity and Corpus Luteum quality of recipients on pregnancy rates following non-surgical transfer of bovine embryos produced in vitro. Oocytes aspirated from small antral follicles of ovaries obtained at a local slaughter house were matured, fertilized with frozen-thawed semen and co-cultured for 6-7 days by utilizing co-culture system with bovine oviduct epithelial cell in vitro. After co-culture, embryos were transfered to recipients on day 7 (estrus=day 0). Recipients were monitored by ultrasonic scanning method or observation for estrus and rectal palpation after 50 days from transfer. The results of this study are follows. 1. Of the 70 recipients, 70%(49 of 70) had not showed estrus sign between day 0 and day 50, but 22.9%(16 of 70) was diagnosed not pregnant. Therefore the overall pregnancy rate of this study was 47.1%(33 of 70). 2. The pregnancy rate of recipients transfered with excellent(66.7%) and good(54.5%) embryos were higher than that of recipients transfered with fair embryos(15.8%) (p<0.05). 3. The pregnancy rate of recipients transfered with morula, compacted morula, blastocyst and expanded blastocysts were 46.2, 55.0, 62.5 and 50.0%, respectively. 4. The pregnancy rates of recipients transfered to heifer and cow were 54.5 and 55.2%, respectively. 5. The pregnancy rates of recipients with CL score I, II(66.7, 63.6%) were higher than those of recipients with CL score III (10%), (p<0.05). Success of transfer of embryos produced in vitro depends on many variables. The important factors identified in this study were the quality of embryos and the CL score of recipient animals after non-surgical transfer of embryos matured, fertilized and cultured in vitro.

• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.207-211
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• 2010

Current developments in IVF and animal cloning have resulted in increasing demand for large quantities of oocytes and ovarian follicles at specific stages of development. These medical and scientific needs may be met by developing an optimal culture system for preantral follicles. In this study, we investigated the growth of porcine preantral follicle cultures in different media and in the presence and absence of serum. Follicles were manually dissected from ovaries obtained from prepubertal gilts at a local slaughterhouse, and cultured for 3 days in M199 or NCSU23 medium supplemented with porcine FSH, transferrin, L-ascorbic acid and insulin. Follicle diameters were measured on day 1 and 3 of culture. In Experiment 1, the effect of supplementing culture medium with fetal calf serum (FCS) on porcine preantral follicle growth was examined. In the group of cultures supplemented with FCS, follicle diameter after 3 days of culture, survival rate and antrum formation rate in the FCS group were significantly higher than those of the control group. In Experiment 2, the effects of culture medium (M199 and NCSU23) on follicle growth were compared. Follicle diameters were increased in the M199 group, compared with those in NCSU23 (p<0.05), but we observed no significant differences in survival and antrum formation rates between cultures grown in the two media. In conclusion, supplementation of the culture medium with serum enhances preantral follicle growth and antrum formation, and M199 is superior to NUSU23 for porcine preantral follicle culture in vitro.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48 no.6
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• pp.438-441
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• 2003

The present study was carried out to assess the possibility of rapid multiplication of Curcuma longa Linne through in vitro culture of shoot-apex. The factor investigated was effect of various growth regulators on shoot-apex culture. The shoot-apex cultured of MS(Murashige and Skoog) medium developed into plantlet in 16 Weeks. M.S. medium containing NAA at 0.5 ppm and BA 5.0 ppm was found to be optimal for growth of in vitro plantlet

• Journal of the Korean Society of Tobacco Science
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• v.13 no.2
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• pp.52-58
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• 1991

Interspecific cross between Nicotiana trigonophylla and N. tabacum cv. BY4 is highly sterile because of abnormal ovule and embryo development. In vitro culture of excised N. trigonophylla ovules after polination by N. tabacum allows significant numbers of hybrid embryos to develop into mature plants. Total yield of seedlings and number of normal seedlings were produced following in vitro culture of individual fertilized ovules of N. trigonophylla X N. tabacum at four days post-pollination on B5 medium containing 6% sucrose. Hybrids were uniform in morphology and peroxidase isozyme composition and the majority were cytologically stable: flower characteristics were generally intermediate between those of the parents. All hybrids evaluated were self-sterile.

• Korean Journal of Veterinary Research
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• v.35 no.1
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• pp.179-185
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• 1995

The fusion rates of nuclear transplant embryos with various DC voltage were 55.6-79.2%. The significantly higher fusion rates of nuclear transplant embryos were achieved at the electric field strenght of DC 1.0-2.0kV/cm(72.0-79.2%) than DC 0.75kV/cm(55.6%, P<0.05). No significant differences in the percentage of embryos that cleaved(48.1, 55.4 and 42.6% respectively) and the percentage of cleaved embryos that developed to morulae/blastocyst(1.9, 5.3 and 1.9% respectively) could be found among the three types of in vitro culture system (Granulosa cell, BOEC co-culture and SOF, P>0.01). The age of the recipient cytoplast(30 vs 40hr) had no effect on the fusion rates and the rates of cleavage development(36.9 vs 44.1%, P>0.01).

• Journal of Embryo Transfer
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• v.5 no.1
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• pp.21-27
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• 1990

The technique for maturation of follicular oocyte has been devised to provide such a low cost and in ptentifut number supply of bovine embryo. Some of problems concerning production of bovine embtyo in vitro were discussed in this paper. Bovine follicular oocytes cultured in vitro achieved normal fertilization but cleavage rates to blastocyst were low compared to the oocyte matured in vivo. It has been concluded that a deficient cytoplasmic maturation occurs in the oocytes matured in vitro. These results indicate that the studies for maturation of bovine follicular oocytes in vitro need improvement of culture conditions and to define the characteristics that might be indicative of healthy oocyte.

• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.394-400
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• 1998

Cryopreservation of embryos by vitrification is a simple method to preserve bovine embryos for subsequent embryo transfer, but embryonic viability after vitrification has been inconsistent and low compared with conventional slow freezing. The aim of the present study is to examine the effect of serum or serum albumin in a vitrification solution and epidermal growth factor(EGF) or fibroblast growth factor(FGF) on in vitro viability of bovine blastocysts frozen by vitrification. Bovine blastocysts were produced by in vitro maturation, fertilization of follicular oocytes and culture of embryos in a synthetic oviduct fluid medium(SOFM) containing BSA and 19 essential and nonessential amino acids. Blastocysts with excellent or good morphology were selected at 7 or 8 days after culture and utilized for vitrification. In experiment 1, blastocysts were vitrified in a solution containing semi-fetal calf serum(SFCS) or BSA(5 or 10mg/ml) and then their subsequent viabilities were examined by culturing thawed embryos in a SOFM containing BSA and 19 amino acids. Effect of EGF or FGF added to a SOFM containing polyvinyl alcohol(PVA) on the viability of vitrified-thawed blastocysts was investigated in experiment 2. BSA added at 5 or 10mg/ml to a vitrification solution showed significantly higher(p < 0.05) developmental rate to expanded and hatching blastocysts than SFCS, but there was no significant difference in the developmental rate to hatched blastocysts after thawing. Supplementation of a culture medium with EGF and/or FGF significantly increased(p < 0.05) embryo development to expanded blastocysts compared with control but showed no beneficial effect on the development to hatching or hatched blastocysts. Coculture of thawed embryos with granulosa cells in a TCM 199 containing 10% fetal calf serum(FCS) showed the highest developmental rate to expanded, hatching and hatched blastocysts among the groups tested. In conclusion, supplementation of a vitrification solution with BSA at 5mg/ml and culture of thawed blastocysts in a medium containing EGF and/or FGF can improve in vitro viability of bovine blastocysts frozen by vitrification.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.1
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• pp.25-32
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• 2010

The present study aimed to examine the effects of ${\gamma}$-linolenic acid (GLA) supplementation to in vitro culture (IVC) medium on in vitro developmental competence, freezability and morphology of in vitro matured and fertilized bovine embryos. In vitro produced (IVP) bovine zygotes were cultured in IVC medium supplemented with 0 (negative control), 15, 31, 62, 125, 250, 500 or 1,000 ppm GLA, 250 ppm linoleic acid albumin (LAA) and without any supplement as a control. Day 6 blastocysts derived from culture control were cultured in IVC medium containing either 62, 250 GLA or 250 LAA for 24 h, and at Day 7 were subjected to freezing or morphological examination by electron microscope. GLA 15 showed a tendency to have a higher cleavage rate at Day 2 (70.3%) than other groups. The hatching rate at Day 9 in LAA (38.2%) was significantly higher than the control and all treatment groups (p<0.05), while the blastocyst rate in LAA (32.4%) did not differ from those of 15 (30.5%), 31 (27.1%), and 62 GLA (33.1%) or the control (35.1%). GLA in concentrations of 125, 250, 500, and 1,000 ppm had significantly detrimental effect on the blastocyst rate compared to 15, 31 and 62 ppm GLA, LAA, and control groups (p<0.05). In contrast, the highest post-thaw survival rate (100%) was observed in the control group (p<0.01). Large lipid droplets were observed in the cytoplasm of trophoblastic cells, even in the control, but were abundant in GLA groups. Taking the results of the study into consideration, the addition of GLA to the culture medium for IVP bovine embryos at the dose of 15 ppm increased the developmental competence of zygotes and enhanced the cleavage rate up to Day 2. However, blastulation rate and post-thaw survival were not increased when GLA was added to the culture media.