• Journal of Embryo Transfer
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• v.33 no.1
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• pp.41-48
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• 2018

In the last several decades, cell therapy research has increased worldwide. Many studies have been conducted on cell therapy, and have revealed that transplanted cells did not survive for long, and implanted cells remained inactive causing immune rejection depending on the patient's condition. Therefore, studies on cell-free therapy need to be conducted. To overcome these limitations, an alternative is the use of supernatant from cells, called "conditioned media (CM)." During in vitro cell culture, culture media supply nutrients to maintain cell characteristics and viability. In the culture, cells not only consume nutrients but also release beneficial proteins and substances, which are called "secretome." CM from cells can be stored for a long time and is easy to handle. Moreover, secretome in CM can also be measured; exact amount of secretome is important to set the standard value for disease treatment. Here, we reviewed studies on CM and confirmed that various secretomes from CM were identified in these studies. Moreover, these findings could benefit cell and animal studies in future. In conclusion, CM could be a potential candidate for an alternative to cell therapy.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.293-301
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• 1996

In the present study, mouse follicular oocytes and 2-cell embryos(late -zygote stage embryos included) were cultured on the electric pad for electric stimulation in the culture incubator. In addition, follicular oocytes and embryos were tested for maturation and development under higher temperature condition($39^{\circ}C$).Mouse follicular oocyte maturation were not affected by anion electric stimulation and there is no significant difference in GBVD and MI between the control and experiment group after 4hr culture. In the embryo culture, it was found that more morula and blastocyst were found in the electric stimulation group rather than the control(96hr). This may seem to be caused with cytoplasmic $Ca^{2+}$ transient rise by electric stimulation(anion pad). On the other hand higher temperature incubation ($39^{\circ}C$) on the anion pad caused all the embryos degenerated within $12h{\sim}24hr$ culture. This was quite different from large animal embryos(bovine, pig, sheep), in which beneficial effect of high temperature incubation for oocyte maturation and embryo development were found.

• Korean Journal of Plant Tissue Culture
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• v.22 no.4
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• pp.207-211
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• 1995

Immature zygotic embryos from immature seeds of Wasabia japonica (cv Dalma) were isolated and cultured on modified MS medium supplemented with 2,4-D, IAA, and BA. Immature zygotic embryos were classified into torpedo shape and cotyledon stage. The highest rates of callus formation were obtained of 1.0mg/L IAA(torpedo stage, 90.0%)and 1.0mg/L 2,4D plus 0.1mg/L BA(cotyledany stage,84.3%). Somatic embryos after 60 days of culture. These numerous somatic embryo could be seperated and subcultured on the same media for further propagation. After 90 days of culture, most somatic embryos were developed well organized embryos which were able to produce into whole plants.

• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.233-238
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• 2000

To obtain a basic information of the development of Genus Viola, morphological and histological observation of in vitro calli and cells in Viola culture cells were investigated. There were two callus types obtained by long term subculture of wild viola (Viola partrinii DC. ) petiole callus. One was friable callus - soft and pale green in color and small cells in size, and the other was compact callus - compact and deep bluish green in color, large cells in size. In scanning electron microscopic observation, friable callus was composed of voculated cell around small. cell clump, while compact callus was composed of cells filled with protoplasm Somatic embryogenesis was observed from suspension culture of the compact callus.

• Asian-Australasian Journal of Animal Sciences
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• v.8 no.6
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• pp.557-562
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• 1995

In this study, characteristics of chick primordial germ cells (PGCs), which is the founder cell of the germline, and gonadal development of the chick embryo between 12hrs and 6 day of incubation were investigated by transverse serial sections of chick embryos under the light microscopic observation. In embryo stage 20 (3 day of incubation), there are a lot of PGCs at the mesenchym, which were moving to the thickened epithelium (gonadal ridge). The PGCs arrive at both right and left gonad primordial in equal number prior to stage 24 (4 day of incubation), but in the following stages, the distribution of the PGCs became asymmetrical. More PGCs colonized the left than the right gonad, but the reason for the unequal distribution of PGCs is uncertain. The PGCs have mostly settled in the gonadal ridge (GR) at 6 day embryo. This study was conducted to investigate characteristics of the PGC migration and gonadal formation and observe the best condition for PGC isolation, culture and to attempt the possibility of the production for transgenic germline chimeras with manipulated PGCs.

• Asian-Australasian Journal of Animal Sciences
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• v.6 no.2
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• pp.319-323
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• 1993

The effect of bovine growth hormone digested with trypsin on whole-body protein synthesis in vitro of chicken embryos was investigated by using a whole-embryo culture system. Bovine growth hormone at 5.3 and 530 ng/ml was digested partially and completely with trypsin for 4 min and 18 h, respectively. After culturing chicken embryos with a synthetic medium containing $L-[4-^3H]$ pheylalanine, whole-embryo protein synthesis was determined from the ratio of specific radioactivities of free and protein-bound pheylalanine. Whole-embryo protein synthesis of the control group cultured with no bovine growth hormone was $49.5{\pm}2.2%/d$. There was no significant interaction between digestion time and the concentration of trypsin-digested bovine growth hormone. Tryptic digestion of bovine growth hormone increased fractional synthesis rates of whole-body protein compared to the 0-min groups, and there was no significant difference between the 4-min and 18-h groups. The higher concentration (530 ng/ml) of trypsin-digested bovine growth hormone was more effective in enhancing whole-embryo protein synthesis than the lower concentration (5.3 ng/ml).

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.8-17
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• 2002

There have been intensive attempts to establish reliable methods far in vitro production (IVP) methods for of porcine embryos. Although a great deal of progress has been made, our current IVP systems still need to be improved. In this review, we focused on studies about in vitro maturation and fertilization (IVM-IVF) of porcine oocytes and their in vitro culture (IVC), especially on an excellent piglets production system using modified IVP system producing porcine blastocysts with high Quality.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2001.05a
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• pp.27-28
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• 2001

• Journal of Animal Science and Technology
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• v.58 no.3
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• pp.11.1-11.6
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• 2016

Background: This study was investigated the effects of culture media conditions on production of eggs fertilized in vitro of embryos from ovaries of high grade Korean native cow, Hanwoo. Methods: The IVMD 101 and IVF 100 were used for in vitro maturation of selected Hanwoo oocytes and In vitro embryo culture after in vitro fertilization, respectively. The IVMD 101 and IVD 101 were used for in vitro culture and completely free of serum. Results: The cleavage rates of 2-cell embryos in reference to Hanwoo oocytes were 86.7, 92.9, and 90.1 % in the control group, IVDM101 medium and IVD101 medium, respectively which indicates that the IVDM101 medium and IVD101 medium may result favorable outcomes. The in vitro development rates of blastocysts were 12.4, 38.4 and 32.4 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. For hatched blastocysts, it was 5.3, 33.9, and 28.6 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. Hence, more favorable results were expected for the hatched blastocysts in which the IVMD101 medium and IVD101 medium were used than the control group. Average cell numbers of blastocysts were 128.3, 165.7, and 163.6 in the groups of TCM-199 + 10 % FBS medium, IVMD 101 medium, and IVD 101 medium, respectively which clearly show that the IVMD 101 and IVD 101 medium consequence significantly higher cell numbers compared to the control group (i.e., TCM-199 +10 % FBS medium). Pregnancy rate after embryo transfer was 39.6 % when the serum free medium was used which is higher than that of the medium supplemented with serum (32.8 %). In addition, stillbirth rates were 4.9 % in the group of serum free medium whereas it was 13.6 % in the serum supplemented medium (13.6 %). Conclusions: Taken altogether, serum free media, the IVMD 101 and IVD 101 represented more favorable results in the embryo development rate of embryos, cell numbers of blastocyst, and pregnancy rate. Of note, the IVMD 101 medium showed better outcomes hence, it might be a better option for future applications for in vitro culture of bovine embryos.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.3
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• pp.419-432
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• 1999

Objective: To establish the evaluation system of the quality of oocytes on the basis of the incidence of cumulus cells apoptosis, to investigate the relationships beween the incidence of cumulus cells and the outcomes of IVF-ET. Method: Thirth-four cycles undergoing controlled ovarian hyperstimulation for IVF-ET with tubal infertility (23 cycles) or unexplained infertility (11 cycles) were included in this study. Cumulus cell masses surrounding mature oocyte and co-culture of embryos with autologous cumulus cells during IVF-ET process. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. The effect of co-culture using cumulus cells and the incidence of cumulus cells apoptosis. Results: The results were as follows: 1. The incidence of apoptosis in cumulus cells markedly increased in patients aged 40 or over, while the fertilization rate was greatly decreased in those age group. 2. Apoptosis in cumulus cells was found in both the fertilized oocytes and unfertilized oocytes, but the incidence of apoptosis was higher in unfertilized oocytes. 3. There is no clear correlation between apoptosis in cumulus cells and the number of oocytes retrieved. However, the incidence of apoptosis was increased when the number of oocytes retrieved was 5 and fewer in comparison with $6{\sim}10$. 4. Embryo grade was significantly affected by the incidence of apoptosis in cumulus cells. 5. Pregnancy rate of IVF-ET per cycle was 29.4%, and the pregnant group had the higher fertilization rate and a significantly lower incidence of apoptosis in cumulus cells compared with the nonpregnant group. 6. When cumulus cells were used as helper cells in the co-culture of the embryo, in vitro activity of cumulus cells based on morphological change and proliferation did not influence the quality of embryo, but was closely associated with the implantation rate and pregnancy rate, which was enhanced when morphological changes and proliferation of cumulus cells was more active. 7. This difference in the outcome of IVF-ET according to in vitro activity of cumulus cells used for co-cultue was not associated with the incidence of apoptosis in cumulus cells; but rather had likely relations with the different secretion pattern of protein, which may be an embryo trophic factor by cumulus cells. Conclusion: These results suggest that the incidence of apoptosis in cumulus cells can be used in predicting oocyte qualities and the outcomes of IVF-ET. And the effect of co-culture largely depends on the in vitro activity of cumulus cells as well.