• The Korean Journal of Physiology and Pharmacology
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• v.24 no.6
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• pp.441-452
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• 2020

In vivo animal models are limited in their ability to mimic the extremely complex systems of the human body, and there is increasing disquiet about the ethics of animal research. Many authorities in different geographical areas are considering implementing a ban on animal testing, including testing for cosmetics and pharmaceuticals. Therefore, there is a need for research into systems that can replicate the responses of laboratory animals and simulate environments similar to the human body in a laboratory. An in vitro two-dimensional cell culture model is widely used, because such a system is relatively inexpensive, easy to implement, and can gather considerable amounts of reference data. However, these models lack a real physiological extracellular environment. Recent advances in stem cell biology, tissue engineering, and microfabrication techniques have facilitated the development of various 3D cell culture models. These include multicellular spheroids, organoids, and organs-on-chips, each of which has its own advantages and limitations. Organoids are organ-specific cell clusters created by aggregating cells derived from pluripotent, adult, and cancer stem cells. Patient-derived organoids can be used as models of human disease in a culture dish. Biomimetic organ chips are models that replicate the physiological and mechanical functions of human organs. Many organoids and organ-on-a-chips have been developed for drug screening and testing, so competition for patents between countries is also intensifying. We analyzed the scientific and technological trends underlying these cutting-edge models, which are developed for use as non-animal models for testing safety and efficacy at the nonclinical stages of drug development.

• Korean Journal of Agricultural Science
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• v.47 no.4
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• pp.979-985
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• 2020

A wide range of techniques have been developed to separate X or Y- chromosome-bearing sperm. In particular, bovine semen sex-sorted by using flow cytometry based on differences in the amount of DNA between X and Y chromosome bearing sperm is used in dairy farms. The first piglets were produced using sex-sorted sperm 30 years ago. However, sexed sperm have not been commercially available in pigs because the flow cytometry technique is not capable of sorting the high number of sperm required for porcine artificial insemination (AI), and the prolonged exposure to an electrical filed might damage to the DNA in sperm. The purpose of this study was to evaluate a boar sperm sorting method based on magnetic nanoparticles. A flow cytometer assay verified the efficacy of the magnetic nanoparticles (> 90% of sex-sorted sperm). In addition, a duplex polymerase chain reaction (PCR) assay using sex chromosome specific genes including SRY (sex-determining region Y; male), ZFY (zinc finger protein Y-linked; male), and ZFX (zinc finger protein X-linked; female) showed that in vitro fertilized porcine embryos by X and Y-chromosome bearing sperm were 100% female (40/40) and 72% female (35/48), respectively, at 8-cell or morula stages, suggesting that the sex-sorted sperm were fertile. In conclusion, our findings suggest that the sex-sorted method based on magnetic nanoparticles can be utilized for porcine sex-sorted AI.

• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.247-256
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• 1999

The present study was performed to improve the reproductive disturbance as well as the elimination of microbiological contamination for animals bred under conventional conditions followed by in vitro fertilization and embryo transfer techniques including embryo and sperm freezing, using a mouse strain(M. m. molossinus-tt＠Kist) showing the abnormal behavior disorder derived from Korean wild mice (Mus musculus molossinus). Moreover, hematological and serum biochemical analyses were also carried out to obtain the basic data of this mouse strain The results are summarized as follows: 1. In comparison with hematological data, the numbers of RBC and platelet of this mouse strain were appeared as the higher value those that of the same aged inbred strains such as BALB/c, DBA/2, C57BL/6 and C3H /Hen. However, no differences were found in values of WBC, Hb and Ht. Moreover, total cholesterol of this strain showed a low value but triglyceride, total protein and albumin values were similar as in inbred strains. 2. The average numbers of superovulated oocytes treated with 2.5/2.5 IU and 5.0/5.0 IU of PMSG/hCG were 11.6 and 12.7, respectively. The fertilization rates of 2.5/2.5 IU PMSG /hCG treatment(87.9%) was higher than 5.0/5.0 IU treatment(52.0%) (p＜0.05) and the developmental rate of 2 cell stage embryos were 외 so appeared as higher value 99.0% and 90.6%, respectively. 3. The rates of in vitro fertilization treated with frozen sperm(24.8%) was significantly lower than of that fresh sperm(87.9%), (p＜0.05). 4. The five, six and ten heads of offspring were obtained from frozen-thawed 2 cell embryos by in vitro fertilized, 2 cell embryos from in vitro fertilized by frozen-thawed spermatozoa. and 2 cell embryos by in vitro fertilization, respectively. These offspring developed the expected disease about 2 weeks after birth, which was confirmed that the disease character of this mutant mouse strain was reliably reproduced. 5. MHV(Mouse hepatitis virus) and Staphylococcus aureus were successfully eliminated from conventional animals by in vitro fertilization-embryo transfer and the use of SPF recipient animals.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.4
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• pp.529-535
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• 2007

The nutritive values of leaves of Citrus grandis, Citrus aurantium, Citrus oranges, Citrus limon, and Citrus deliciosa were evaluated by chemical composition and in vitro gas production techniques. There were significant (p<0.001) differences among citrus species in terms of chemical composition. Crude protein (CP) contents ranged from 123.0 to 148.3 g/kg DM. Neutral detergent fibre (NDF) and acid detergent fibre (ADF) contents were varied with species in the range 219.4-355.4 and 215.0-278.8 g/kg DM respectively. Condensed tannin (CT) contents were ranged from 5.9 to 10.2 g/kg DM. The PEG addition significantly (p<0.001) increased the gas production and some estimated parameters of citrus tree leaves. However, species showed variable responses to polyethylene glycol (PEG) treatment. There were also significant (p<0.001) differences among species in terms of gas production and estimated parameters. The OMD and ME contents of citrus leaves without PEG supplementation were ranged from 66.5 to 73.3% and 9.8 to 10.9 MJ/kg DM respectively. The improvement in gas production, organic matter digestibility (OMD) and metabolizable energy (ME) with PEG emphasized the negative effect of tannins on digestibility. The increase (%) in the estimated OMD and ME contents ranged from 5.5 to 9.8% and 5.7 to 10.2% respectively. All citrus tree leaves studied in this experiment have potential nutritive values indicated by high crude protein content, OMD, ME and low fiber values.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.4
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• pp.496-500
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• 2007

Holstein cow ovaries obtained at a slaughterhouse were used to study the influence of the oocyte collection methods (slicing, puncture, aspiration I and II) on recovery efficiency and subsequent in vitro maturation and embryonic development competence of immature oocytes recovered. In the slicing method, the whole ovarian was chopped into small pieces with a surgical blade. In the puncture method, the whole ovarian surface was punctured by 18-g needle. In other 2 aspiration methods, collected oocytes by aspirating from the visible follicles using an 18-g needle attached to a 5 ml syringe (aspiration I) or using a constant negetive pressure (-80 mmHg) with a vacuum pump (aspiration II). The oocytes were classified into 4 classes on the basis of the morphology of cumulus cells and cytoplasmic appearance of oocyte. Slicing ($9.6{\pm}0.4$) and puncture ($9.7{\pm}0.4$)yielded a larger number of oocytes per ovary than other two aspiration methods (aspiration I and II were $5.8{\pm}0.3$and $5.6{\pm}0.4$, respectively) (p<0.05). The number of the highest quality oocytes (grade A) per ovary was significantly higher in slicing ($4.2{\pm}0.2$) and puncture ($4.6{\pm}0.1$) methods than in other methods (aspiration I and II were $1.2{\pm}0.2$ and $1.4{\pm}0.2$, respectively) (p<0.05). The rate of nuclear maturation of the highest and higher quality oocytes (grade A and grade B, respectively) was not affected by the oocytes collection methods. The oocytes collection methods also did not influence subsequent embryonic developmental competence after in vitro fertilization with M II stage oocytes. It is concluded that slicing and puncture methods of the ovaries can be used as an alternative techniques to aspiration by the syringe or vacuum pump.

• The Journal of Advanced Prosthodontics
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• v.4 no.3
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• pp.162-169
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• 2012

PURPOSE. Fracture of the veneering material of zirconia restorations frequently occurs in clinical situations. The purpose of this in vitro study was to compare the fracture strengths of zirconia crowns veneered with various ceramic materials by various techniques. MATERIALS AND METHODS. A 1.2 mm, $360^{\circ}$ chamfer preparation and occlusal reduction of 2 mm were performed on a first mandibular molar, and 45 model dies were fabricated in a titanium alloy by CAD/CAM system. Forty-five zirconia copings were fabricated and divided into three groups. In the first group (LT) zirconia copings were veneered with feldspathic porcelain by the layering technique. In the second group (HT) the glass ceramic was heat-pressed on the zirconia coping, and for the third group (ST) a CAD/CAM-fabricated high-strength anatomically shaped veneering cap was sintered onto the zirconia coping. All crowns were cemented onto their titanium dies with Rely $X^{TM}$ Unicem (3M ESPE) and loaded with a universal testing machine (Instron 5583) until failure. The mean fracture values were compared by an one-way ANOVA and a multiple comparison post-hoc test (${\alpha}$= 0.05). Scanning electron microscope was used to investigate the fractured interface. RESULTS. Mean fracture load and standard deviation was $4263.8{\pm}1110.8$ N for Group LT, $5070.8{\pm}1016.4$ for Group HT and $6242.0{\pm}1759.5$ N for Group ST. The values of Group ST were significantly higher than those of the other groups. CONCLUSION. Zirconia crowns veneered with CAD/CAM generated glass ceramics by the sintering technique are superior to those veneered with feldspathic porcelain by the layering technique or veneered with glass ceramics by the heat-pressing technique in terms of fracture strength.

• Advances in nano research
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• v.7 no.4
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• pp.241-248
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• 2019

Recent researches demonstrated well promising anticancer activities for antibiotics. Such effects would be significantly increased while nanoparticle based delivery systems were applied. In this study, the goal was aim to improve anticancer and antitoxic effects of Streptomycin by loading on special kind of dendrimer (anionic-linear-globular second generation). In the current study, Size and zeta potential as well as AFM techniques have been used to prove the fact that the loading was performed correctly. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the drug loaded on dendrimer nanoparticle were determined and compared with both of dendrimer alone and free drug with respect to staphylococcus aureus as the test microorganism. The anticancer activity among three groups including Streptomycin, Streptomycin -G2 dendrimer, and control was measured in vitro. In vitro studies showed that G2 anionic linear-globular polyethylene-glycol-based dendrimer, which loaded on Streptomycin was able to significantly improve the treatment efficacy over clinical Streptomycin alone with respect to proliferation assay. Maximal inhibitory concentration (IC50) was calculated to be $257{\mu}g/mL$ for streptomycin alone and $55{\mu}g/mL$ for Streptomycin -G2 dendrimer. In addition, Streptomycin -G2 dendrimer conjugate prevented the growth of MCF-7 cancerous cells in addition to enhance the number of apoptotic and necrotic cells as demonstrated by an annexin V-fluorescein isothiocyanate assay. Streptomycin -G2 dendrimer conjugate was able to increase Bcl-2/Bax ratio in a large scale compared with the control group and Streptomycin alone. Based on results a new drug formulation based nano-particulate was improved against S. aureus with sustained release and enhanced antibacterial activity as well as anticancer activity shown for functional cancer treatment with low side effects.

• Journal of Biomedical Engineering Research
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• v.22 no.5
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• pp.413-418
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• 2001

We developed shunt valves used to treat patients with hydrocephalus. The valves under development were constant Pressure type ventriculoperitoneal (VP) calves made of silicone elastomer. In vitro experiments showed that our valves had similar Pressure-flow control characteristics to the valved currently available in the market. Our valves also showed competent performance in the 28 days of continuous pumping tests acording to the IS07197 specifications. We artificially inducted hydrocephalus to a 10kg beagle do9. The size of the ventricles of the dog was substantially increased and the dog showed abnormal behavior. After our valve being implanted, the ventricles recovered regular size with the normal behavior observed in the dog. The flow orifice of the shunt valve diaphragm was in the older of 10$\mu$m during calve operation and hence the pressure-flow control characteristics tended to change by a small chance in the valve dimension. Therefore, precision design and manufacturing techniques were necessary for successful operations of the shunt valves .

• Journal of Embryo Transfer
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• v.20 no.1
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• pp.9-15
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• 2005

This study was conducted to investigate the effects of various factors of practitioner's techniques such as skill, difficulty, consumed time, implantation depth, bleeding and altering of implant location, on pregnancy and abortion after transfer of in vitro produced Korean Native Cow embryos. The pregnancy rate of skilled transfer $(54.2\%)$ was significantly higher than that of unskilled transfer $(37.9\%)$, of 2/3 $(46.9\%)$ location in uterus was significantly higher than in 1/2 $(39.5\%)$ or 4/5 $(38.0\%)$ uterus, and of altering implantation location $(52.9\%)$ was significantly increased. There were no difference in pregnancy among the groups of difficulty of transfer, consumed time and bleeding of uterus. The abortion rates from skill, difficulty of transfer, implantation depth, consumed time, bleeding of uterus and altering of location were not differ.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2006.11a
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• pp.43-58
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• 2006

As a bioreactor, bird has proved to be most efficient system for producing useful therapeutic proteins. More than half of the egg white protein content derives from the ovalbumin gene with four other proteins(lysozyme, ovomucoid, ovomucin and conalbumin) present at levels of 50 milligrams or greater. And the naturally sterile egg also contains egg white protein at high concentration allowing for a long shelf life of recombinant protein without loss in activity. In spite of these advantages, transgenic procedures for the bird have lagged far behind because of its complex process of fertilized egg and developmental differences. Recently, a system to transplant mouse testis cells from a fertile donor male to the seminiferous tubules of an infertile recipient male has been developed. Spermatogenesis is generated from transplanted cells, and recipients are capable of transmitting the donor haplotype to progeny. After transplantation, primitive donor spermatogonia migrate to the basement membrane of recipient seminiferous tubules and begin proliferating. Eventually, these cells establish stable colonies with a characteristic appearance, which expands and produces differentiating germ cells, including mature spermatozoa. Thus, the transplanted cells self-renew and produce progeny that differentiate into fully functional spermatozoa. In this study, to develop an alternative system of germline chimera production that operates via the testes rather than through developing embryos, the spermatogonial stem cell techniques were applied. This system consisted of isolation and in vitro-culture of chicken testicular cells, transfer of in vitro-maintained cells into heterologous testes, production of germline chimeras and confirmation of germline transmission for evaluating production of heterologous, functional spermatozoa.