• 한국수정란이식학회지
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• 제5권1호
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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.

• 한국가축번식학회지
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• 제12권2호
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• pp.112-119
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• 1988

These studies were conducted to investigate the effects of culture temperature and time on the in vitro maturation and semen type and media on the in vitro fertilization of bovine follicular oocytes, and to asses in vitro fertilization rate of oocytes cultured by extraffollicular method following fertilization in vitro, or transfer into the pseudopregnant rabbit oviduct or uterus. The bovine oocytes recovered from follicles were cultrued for 18 hrs or 72hrs at 38$^{\circ}C$ with 5% CO2 in moist air. Flesh-diluted(2 folds) and frozen-thawed semen in 0.5ml straw from a fertile bull were used. In order to obtain capacitation of spermatozoa were treated with bovine follicular fluids(BFF) and Inophore A(IA). The results obtained were summarized as follows: 1. The oocytes were classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 62.0%, 12.0%, 17.2%, 5.9% and 3.0% of the total oocytes harvested, respectively. 2. The oocytes matured to metaphase II were significantly increased between 24-48hrs of incubation and at 37-39$^{\circ}C$ with 5% CO2 in moist air. 3. The in vitro fertilization rate following transferred into rabbit oviduct or uterus with bull semen and in vitro matured oocytes were higher ligation than non-ligation of oviduct or uterus. 4. The in vitro fertilization rate of oocytes matured in vitro were higher neat than frozen semen and treatment of IA than BFF on the capacitation of spermatozoa. 5. The effects of semen types and media on in vitro fertilization of oocytes matured in vitro were higher fertilization rate of neat than friozen semen, and media was not significant.

• Restorative Dentistry and Endodontics
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• 제46권2호
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• pp.19.1-19.12
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• 2021

Objectives: The purpose of this study was to conduct a systematic review and meta-analysis of in vitro studies regarding the effectiveness of reciprocating and rotary instrumentation on microbial reduction in root canals. Materials and Methods: PubMed, Scopus, Web of Science, the Cochrane Library, and the gray literature were searched through December 2019. Studies comparing the influence of reciprocating and rotary instrumentation on the removal of microorganisms from root canals that quantified the antimicrobial effect were included. Data extraction was completed using a systematic form for data collection. The risk of bias of the studies was evaluated. Standardized mean differences (SMDs) and confidence intervals (CIs) were calculated using a random effects meta-analysis. Results: Seventeen in vitro studies were included in this systematic review, of which 7 provided adequate data for inclusion in the meta-analysis. Both reciprocating and rotary systems were similarly effective in reducing the microbial load in infected root canals (SMD [95% CI], 0.0481 [-0.271, 0.367]). Three studies showed a low risk of bias, whereas most of the studies (82%) presented a medium risk. Conclusions: Although both techniques decrease the microbial content (with reductions of 23.32%-88.47% and 23.33%-89.86% for reciprocating and rotary instrumentation, respectively), they are not able to provide complete disinfection of root canals.

• 대한화장품학회지
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• 제28권3호
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• pp.171-184
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• 2002

인체실험(in vivo) 방법과의 상관성이 우수한 in vitro 자외선차단지수(SPF) 측정방법을 개발하고자 본 연구를 수행하였다. 표준자외선차단제인 8% homomentyl salicyate(HMS) 제제와 P3 reference standard 제제 및 기능성화장품중 자외선차단화장품을 사용하여 자외선차단지수측정기(SPF 290 analyzer)를 가지고 in vitro 자외선차단지수를 측정하고, 인체실험과의 상관계수를 구하였다. 8% HMS 제제 및 P3 Reference standard 제제의 in vitro SPF 결과는 식약청고시 2001-64호의 규정과 유사한 결과를 나타내었고, 크림 및 로오숀 제형에서는 상관계수 0.9506 및 0.9769로서 인체실험과 높은 상관성을 나타내었다. 메이크업베이스 및 리퀴드파운데이션, 사용시 흔들어 쓰는 로오숀, 압축분말은 각각 상관계수 0.8812, 0.8632, 0.5984를 나타내었으며. 압축분말의 도포를 위해서는 크림베이스와 1:0.8의 비율로 섞는 것이 가장 좋은 결과를 나타내었다 본 실험결과를 통해 사람을 이용한 자외선차단지수 측정방법을 대체할 수 있을 것으로 기대되며, 이 후 자외선차단 화장품의 품질관리에 적용하고, 고시개정의 기초자료로 활용될 수 있을 것으로 기대된다.

• 한국가축번식학회지
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• 제14권1호
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• pp.73-83
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• 1990

These studies were carried out to find the proper conditions for in vitro maturation and fertilization of bovine follicular oocytes and culture methods capable of further developing early embryos. For these objectives, the cleavage rate of oocytes matured and fertilized in vitro was investigated under medium supplemented with hormones and estrous cow serum and season of oocytes collection as well as different cumulus cell stage before insemination. Finally, 2~8 cell embryos were cultured in in vitro and in vitro culture system to investigate developmental capacity into morula. 1. Cleavage rate of oocytes matured in vitro was 27%(20/73) for A(LH＋FSH＋estradil-17$\beta$＋10% FBS), 38%(27/71) for B(LH＋10% ECS) and 27%(15/56) for C(10% ECS), respectively. Supplement B showed more higher rate and 4~8 cell embryos were also obtained much more in this group(67%, 18/27). In vitro maturation rate of follicular oocytes cultured in TCM 199 supplemented withLH and 10% ECS was 88%(75/85). 2. Cleavage rate(15%, 10/65) of oocytes collected in summer was significantly lower than in fall(47%, 42/89). 3. Cleavage rate(15%, 10/65) of oocytes collected in summer was significantly lower than in fall(47%, 42/89). 3. Cleavage rate(15%, 10/65) of oocytes with partially removed cumulus cell before insemination was more higher than that(44%, 27/62) of oocytes with intact cumulus cell. 4. The frequency of development from early cleaved embryos into morula was 6%(4/65), 12%(4/33) for co-culture of cumulus cell monolayer and bovine oviduct epithelial cell monolayer, respectively and 25%(6/25) in ligated rabbit oviduct.

• Journal of Pharmaceutical Investigation
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• 제38권3호
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• pp.183-189
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• 2008

The present study compared the feasibility of Caco-2 and MDCK cells as an efficient in-vitro model for the drug classification based on Biopharmaceutics Classification System (BCS) as well as an in-vitro model for drug interactions mediated by P-gp inhibition or P-gp induction. Thirteen model drugs were selected to cover BCS Class I{\sim}IV$ and their membrane permeability values were evaluated in both Caco-2 and MDCK cells. P-gp inhibition studies were conducted by using vinblastine and verapamil in MDCK cells. P-gp induction studies were also performed in MDCK cells using rifampin and the P-gp expression level was determined by western blot analysis. Compared to Caco-2 cells, MDCK cells required shorter period of time to culture cells before running the transport study. Both Caco-2 and MDCK cells exhibited the same rank order relationship between in-vitro permeability values and human permeability values of all tested model compounds, implying that those in-vitro models may be useful in the prediction of human permeability (rank order) of new chemical entities at the early drug discovery stage. However, in the case of BCS drug classification, Caco-2 cells appeared to be more suitable than MDCK cells. P-gp induction by rifampin was negligible in MDCK-cells while MDCK cells appeared to be feasible for P-gp inhibition studies. Taken all together, the present study suggests that Caco-2 cells might be more applicable to the BCS drug classification than MDCK-cells, although MDCK cells may provide some advantage in terms of capacity and speed in early ADME screening process.

• 한국동물생명공학회지
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• 제37권4호
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• pp.285-291
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• 2022

In 1951, Colin Russell Austin and Min Chueh Chang identified "capacitation", a special process involving ejaculated spermatozoa in the female reproductive tract. Capacitation is a phenomenon that occurs in vivo, but almost all knowledge of capacitation has been obtained from in vitro studies. Therefore, numerous trials have been performed to establish in vitro capacitation methods for various studies on reproduction. Although a series of studies have been conducted to develop an optimal protocol for inducing capacitation, most have focused on identifying the appropriate chemical compounds to induce the capacitation of boar spermatozoa in vitro. Therefore, the purpose of this study was to identify the optimal incubation time for inducing capacitation in vitro. Duroc semen was incubated for various periods (60, 90, and 120 min) to induce capacitation. Sperm function (sperm motility, motion kinematic parameters, and capacitation status) was evaluated. The results showed that total sperm motility, rapid sperm motility, progressive sperm motility, curvilinear velocity, and average path velocity significantly decreased in a time-dependent manner. However, the capacitation status did not show any significant changes. Taken together, these results indicate that an incubation time of more than 60 min suppresses sperm motility and motion kinematic parameters. Therefore, we suggest that 60 min may be the best incubation time to induce capacitation without negative effects on sperm motility and motion kinematics in boar spermatozoa in vitro.

• 한국가축번식학회지
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• 제16권3호
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• pp.209-215
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• 1992

These studies were carried out to overcome 2-cell block and in vitro development to blastocysts in vitro fertilization of mouse embryos. The unfertilized ova were obtained by superovulation in ICR mice of 4 to 6 weeks old. Tyrode's 280 solution was used as basal media, and the pH range of media examined was designed from 6.5 to 7.5 with 0.2 interval and the range of osmolality from 250 to 370 mOsm with 20 interval, and the period of sperm preincubation examined was 30, 60, 120, and 180 minutes. The ova developed to 2-cell embryos after 26hrs of incubation with preincubated sperm were evaluatated as in vitro fertilized ones. The results obtained were summarized as follows: 1. The optimal ranges of pH and osmolality of culture media and of sperm preincubation time for in vitro development of in vitro fertilized ova to blastocyst were pH 7.1 to 7.3, 250 to 350 mosmol and 60 to 180 min, respectively. 2. With the media of pH 7.1, 310 mOsm and sperm preincubation period of 120min in another experiment of large sample size, the in vitro fertilized ova was found 66.5% and the in vitro development of in vitro fertilized ova to blastocyst was found 35.8%. From the above results it was concluded that the optimal conditions of pH and osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOam and 120min, respectively.

• 한국가축번식학회지
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• 제21권3호
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• pp.275-280
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• 1997

This study was carried out to investigate on the survival and in vitro developmental rates of bisected bovine embryos by microblade, micropipette and pronase methods. Bisected embryos cultured for 1∼7 days in TCM-199 media with 10 FCS＋hormones. Survival and in vitro developmental rates was defined on in vitro culture or FDA-test. The results are summarized as follows ; 1. The survival and in vitro developmental rates of bisected bovine embryos by microblade, micropipette and pronase methods were 22.2, 16.7, 15.0% and 22.2, 23.3, 18.8%, respectively. In vitro developmental rate of bisected bovine embryos was significantly lower than that of non-bisection embryos(27.8% and 25.0%). 2. In vitro developmental rates of bovine embryos bisected for 1, 2, 4, 8, 16 cells stages during in vitro culture in 10% FCS＋TCM-199 media were 25.0, 20.0, 20.0, 15.0 and 6.7%, respectively. 3. In vitro developmental rates of intact and free-zona pellucida of bisected demi-embryos during in vitro culture in 10% FCS＋TCM-199 media were 25.6, 16.7%, respectively. 4. In vitro developmental rates of biopsied embryos and biopsied blastomeres during in vitro culture in 10% FCS＋TCM-199 media were 20.0, 11.1%, respectively.

• 한국수정란이식학회지
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• 제29권1호
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• pp.1-5
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• 2014

Recent 2 decades, including in vitro maturation (IVM), assisted reproductive technologies (ARTs) achieved noteworthy development. However the efficiency of ARTs with in vitro matured oocytes is still lower than that with in vivo oocytes. To overcome those limitations, many researchers attempted to adapt co-culture system during IVM and consequently maturation efficiency has been increased. The beneficial effects of applying co-culture system is contemplated base on communication and interaction between various somatic cells and oocytes, achievement of paracrine factors, and spatial effects of extracellular matrix (ECM) from somatic cell surface. The understanding of co-culture system can provide some information to narrow the gap between in vitro and in vivo. Here we will review current studies about issues for understanding cu-culture system with various somatic cells to improve in vitro maturation microenvironment and provide bird view and strategies for further studies.