• Title/Summary/Keyword: In-vitro techniques

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Neuron-on-a-Chip technology: Microelectrode Array System and Neuronal Patterning (뉴런온칩 기술: 미세전극칩시스템과 신경세포 패터닝 기술)

  • Nam, Yoon-Key
    • Journal of Biomedical Engineering Research
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    • v.30 no.2
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    • pp.103-112
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    • 2009
  • Neuron-on-a-Chip technology is based on advanced neuronal culture technique, surface micropatterning, microelectrode array technology, and multi-dimensional data analysis techniques. The combination of these techniques allowed us to design and analyze live biological neural networks in vitro using real neurons. In this review article, two underlying technologies are reviewed: Microelectrode array technology and Neuronal patterning technology. There are new opportunities in the fusion of these technologies to apply them in neurobiology, neuroscience, neural prostheses, and cell-based biosensor areas.

Journal Review and Some Clinical Guidelines on Resin-Bonded Fixed Prosthodontics (Resin-bonded Prosthesis에 대한 문헌 고찰과 성공을 위한 제언)

  • Cho, Lee-Ra
    • Journal of the Korean Academy of Esthetic Dentistry
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    • v.10 no.1
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    • pp.56-70
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    • 2001
  • The use of resin-bonded fixed partial dentures described in the early 1980s caused an conservative way to preserve tooth structure in the restorative dental community. The treatment of patients with requires long term analysis of clinical application and basic research. Failure rates of these prosthesis ranged from 3% to 55%. These varieties were orginated by different techniques, materials, tooth preparation methods and diverse clinical situations. This article review was focused on the standard long term results and in vitro studies on bond strength between metal and teeth. From this, many useful clinical guidelines to RBFPD could be adopted to clinical dentistry. For successful results, careful case selection and good clinical skills are needed. And appropriate techniques for each situations should be adopted. Also, RBFPD using new materials like all-ceramics, FRC/Ceromer was introduced.

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Reproductive Biotechnologies for Improvement of Buffalo: The Current Status

  • Purohit, G.N.;Duggal, G.P.;Dadarwal, D.;Kumar, Dinesh;Yadav, R.C.;Vyas, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1071-1086
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    • 2003
  • Reproductive biotechnologies continue to be developed for genetic improvement of both river and swamp buffalo. Although artificial insemination using frozen semen emerged some decades back, there are still considerable limitations. The major problem appears to be the lack of efficient methods for estrus detection and timely insemination. Controlled breeding experiments in the buffalo had been limited and similar to those applied in cattle. Studies on multiple ovulation and embryo transfer are essentially a replica of those in cattle, however with inherent problems such as lower number of primordial follicles on the buffalo ovary, poor fertility and seasonality of reproduction, lower population of antral follicles at all stages of the estrous cycle, poor endocrine status and a high incidence of deep atresia in ovarian follicles, the response in terms of transferable embryo recovery has remained low with 0.51 to 3.0 per donor and pregnancy rates between 15 to 30%. In vitro production of buffalo embryos is a valid alternative to recovery of embryos by superovulation. This aspect received considerable attention during the past decade, however the proportion of embryos that develops to the blastocyst stage is still around 25-30% and hence the in vitro culture procedures need substantial improvement. Embryo cryopreservation procedures for direct transfer post thaw need to be developed for bubaline embryos. Nuclear transfer and embryo cloning is a technique that has received attention in various species during recent years and can be of immense value in buffaloes as they have a low rate of embryo recoveries by both in vitro and in vivo procedures. Gender pre-selection, genome analysis, gene mapping and gene transfer are a few of the techniques that have been studied to a limited extent during recent years and are likely to be included in future studies on buffaloes. Very recently, reproductive biotechnologies have been applied to feral buffaloes as well, but the results obtained so far are modest. When fully exploited they can play an important role in the preservation of endangered species.

Development of Production Techniques for Korean Native Cattle Calves from Embryos by In Vitro Technology 3. Effects of Culture Medium on Blastocyst Development and Effects of Sucrose and Trehalose on the Survival Rate of In Vitro Developed Embryos after Thawing (체외배양기술로 생산된 초기배에 의한 한우 송아지 생산기술 III. 배반포와 발생에 미치는 발생배지의 효과와 동결융해후의 생존율에 미치는 Sucrose와 Trehalose의 효과)

  • 서경덕;김호중;김갑수;김광식
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.173-178
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    • 1998
  • The objective of this study was to evaluate the embryonic development ability and the appearance of blastocysts of bovine in vitro fertilized oocytes cultured in different culture media, and also to evaluate survival rate after thawing of frozen embryos by using 1.5 or 1.8M ethylene glycol(EG) with sucrose or trehalose. Fertilized oocytes were divided into three groups; i ) monolayer of cumulus /granulosa cell prepared by TGM 199+5% calf serum(TGM199), ii)GRlaa+5% CS, iii)SOF+5% CS, and they were cultured after insemination for 9 days, at 39˚C, under 5% $CO_2$ in air, but SOF+5% CS was cultured at 39˚C, under 5% 02, 5% GO2, 99% N2. Blastocysts derived from GRlaa + 5% CS on day 7~8 after insemination were frozen by using 1.5M EG or 1.8M EG with/without 0.2M sucrose or O.1M trehalose. The development rate of blastocysts on day 7 after insemination in SOF+5% CS was significant higher than in TCM199 or CR1aa(P<0.05). The appearance rate of blastocysts on day 7-8 after insemination was higher than in TCM199, when fertilized oocytes were cultured in GRlas or SOF. The survival rate of frozen blastocysts after thawing tended to increase, when blastocysts were frozen by using 1.8M EG with 0.2M sucrose or O.1M trehalose. These results indicated that SOF or CRlaa media with amino acids was superior to TCM199 with monolayer in terms of blastocyst development in culturing of in vitro fertilized bovine nocytes, and sucrose or trehalose was supposed to prevent embryos from the freezing shock.

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Evaluation of porcine intestinal organoids as an in vitro model for mammalian orthoreovirus 3 infection

  • Se-A Lee;Hye Jeong Lee;Na-Yeon Gu;Yu-Ri Park;Eun-Ju Kim;Seok-Jin Kang;Bang-Hun Hyun;Dong-Kun Yang
    • Journal of Veterinary Science
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    • v.24 no.4
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    • pp.53.1-53.12
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    • 2023
  • Background: Mammalian orthoreovirus type 3 (MRV3), which is responsible for gastroenteritis in many mammalian species including pigs, has been isolated from piglets with severe diarrhea. However, the use of pig-derived cells as an infection model for swine-MRV3 has rarely been studied. Objectives: This study aims to establish porcine intestinal organoids (PIOs) and examine their susceptibility as an in vitro model for intestinal MRV3 infection. Methods: PIOs were isolated and established from the jejunum of a miniature pig. Established PIOs were characterized using polymerase chain reaction (PCR) and immunofluorescence assays (IFAs) to confirm the expression of small intestine-specific genes and proteins, such as Lgr5, LYZI, Mucin-2, ChgA, and Villin. The monolayered PIOs and three-dimensional (3D) PIOs, obtained through their distribution to expose the apical surface, were infected with MRV3 for 2 h, washed with Dulbecco's phosphate-buffered saline, and observed. Viral infection was confirmed using PCR and IFA. We performed quantitative real-time reverse transcription-PCR to assess changes in viral copy numbers and gene expressions linked to intestinal epithelial genes and antiviral activity. Results: The established PIOs have molecular characteristics of intestinal organoids. Infected PIOs showed delayed proliferation with disruption of structures. In addition, infection with MRV3 altered the gene expression linked to intestinal epithelial cells and antiviral activity, and these effects were observed in both 2D and 3D models. Furthermore, viral copy numbers in the supernatant of both models increased in a time-dependent manner. Conclusions: We suggest that PIOs can be an in vitro model to study the infection mechanism of MRV3 in detail, facilitating pharmaceutical development.

Study on Production of Cloned Animals by Recycling Nuclear Transplantation I. Activation of Nuclear Recipient Oocytes by Electrostimulation in Rabbits (반복핵이식에 의한 복제동물 생산에 관한 연구 I. 토끼 수핵난자의 전기자극에 의한 활성화)

  • 이효종;최민철;최상용;박충생;윤창현;강대진
    • Journal of Embryo Transfer
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    • v.8 no.2
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    • pp.151-157
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    • 1993
  • The present study was undertaken to determine the optimal condition for parthenogenetic activation of rabbit oocytes by electric stimulation in vitro in an attempt to develop nuclear transplantation techniques for cloning mammalian embryos and animals. Freshly ovulated oocytes were collected from superovulated rabbits from 13 to 26 hrs. after hCG injection. The cumulus-free oocytes were activated parthenogetically by repeated stimuli of square direct electric pulses in O.3M mannitol solution. After applying electric stimulations of different voltages, pulse durations and pulse times, all of the oocytes were cultured in TCM-199 with 10% FCS for 96 hours in a 5% $CO_2$ incubator, and their developmental potential in vitro was examined. The higher activation rate (68.9%) was achieved at the voltage of 2.0kv/cm, the pulse duration of 60 $\mu$sec and three pulse times and the activation rate of 100% was achieved at the pulse duration of 100 and 200 $\mu$sec, the voltage of 1.5kv/cm and three pulse times. Although the higher rates of activation of oocytes were achieved at 100 and 200 $\mu$sec, none of them developed to blastocyst in vitro. The oocytes collected 18~20 hours post hCG injection showed the highest rate of activation and development to blastocyst in vitro than the oocytes collected 13~15 or 25~26 hours post hCG injection. Therefore, it can be suggested that the application of electric stimulation of 2.0kv/cm, 60 $\mu$sec and three pulse times to the oocytes collected at 18~20 hours post hCG injection would be more beneficial for the parthenogenetic activation of oocytes in rabbits.

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DETERMINATION OF THE APPARENT ILEAL DIGESTIBILITY OF PROTEIN AND AMINO ACIDS IN FEEDSTUFFS AND MIXED DIETS FOR GROWIG-FINSHING PIGS WITH THE MOBILE NYLON BAG TECHNIQUE

  • Yin, Y.L.;Zhong, H.Y.;Huang, R.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.5
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    • pp.433-441
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    • 1995
  • A series of experiments was conducted to determine the influence of various pepsin-HCL pretreatment factor, hereby the factors of duration of washing for the retrieved bags, inherent to the mobile nylon bag technique (MNBT), on apparent ileal digestibility of crude protein (AIDCP) and apparent ileal digestibility of dry matter (AIDDM). At last, the AIDCP and apparent ileal digestibility of amino acids (AIDAA) in maize, barley, wheat, rapeseed meal, cottonseed meal and three mixed diets were determined with the MNBT and ileo-rectal anastomis pigs (IRAT). For the MNBT techniques, bag measuring $25{\times}40$ MM and containing 0.75 g feedstuff samples, after pre-digestion in vitro, were introduced into the ileo-rectal anastomis pigs (IRAT) gastrointestinal tract through a duodenal cannula and recovered in the ileal digesta between 6 and 12 h. later. 1. The apparent ileal digestibility of dry matter (AIDDM) and crude protein (AIDCP) of the tested samples, with the exception of fish meal, determined by MNBT were not affected by the different pepsin-HCL pretreatment times in vitro between 2.5 h. and 4 h. 2. There was no significant (p > 0.05) difference of the AIDCP and AIDDM of maize determined by the MNBT among different pepsin concentration (0.03%, 0.07% and 0.1 %) treatment in vitro. 3. The AIDCP determined with the MNBT was affected by the washed and unwashed recovered bags from the ileal digesta. 4. The AIDCP and AID amino acids (AIDAA) of maize, barley, wheat, rapeseed meal, soya-bean meal, cottonseed meal and three mixed diets from the MNBT, with a solution of 0.01N HCL (PH 2) and 0.1% of pepsin concentration, a pepsin-HCL pretreatment time in vitro or 4h. and a washing time of the recovered bag from the ileal digesta compared well with those from the IRAT. The linear regression analysis showed a significant correlation (p < 0.01) of AIDCP and AIDDA between the IRAT and MNBT.

A Case of Successful Pregnancy with Electroejaculation and In Vitro Fertilization (전기사정과 시험관아기시술에 의한 성공적인 임신 1례)

  • Nam, Y.S.;Kim, H.J.;Jun, Y.J.;Kim, H.K.;Oum, K.B.;Yoon, T.K.;Cha, K.Y.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.117-121
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    • 1999
  • Although anejaculation is a relatively uncommon occurrence in the general population, over 12,000 new cases are reported annually. Anejaculation may result from spinal cord injury, retroperitoneal lymph node dissection, diabetes mellitus, transverse myelitis, multiple sclerosis, or psychogenic disorders. At least 30% of men with this problem are or will be married and many will seek help to remedy their infertile state. The evolution of technique and instrumentation over the last 30 years has made electroejaculation an accessible and acceptable form of therapy. Recent successes in inducing ejaculation by means of rectal probe electrostimulation or vibratory stimulation combined with assisted reproductive techniques, such as zygote intrafallopian transfer (ZIFT), gamete intrafallopian transfer (GIFT), and in vitro fertilization (IVF), have provided these men means of producing their own biologic offspring. We have experienced a successful pregnancy with electroejaculation and in vitro fertilization in a infertile patient whose husband had an ejaculatory disturbance due to a spinal cord injury. So we report this case with a brief review of literatures.

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Effect of Medium Composition on in Vitro Shoot Regeneration from Leaves of Cassava (Manihot esculenta Crantz) Through Somatic Embryogenesis and Callus Induction (카사바 잎 절편 유래 체세포배 배양시 배지조성이 기내 식물체 재분화에 미치는 영향)

  • Young Hee Kwon;Joung Kwan Lee;Hee Kyu Kim;Kyung Ok Kim;Ju Hyoung Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.19-19
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    • 2020
  • The Cassava(Manihot esculenta Crantz) is a tropical root crop, originally from Amazonia, that provides the staple food of an estimated 800 million people worldwide. It belongs to the family Euphorbiaceae which also includes rubber (Hevea brasiliensis) and castor bean (Ricinus communis). Among tropical crops, rice, sugarcane, maize and cassava are the most important sources of calories for human consumption. Problems in the propagation of cassava are virus diseases and low rates of seed germination. So we tried to optimize protocols for mass production of somatic embryo amenable to large-scale vegetative propagation of Cassava. After in vitro eight-week culture of leaves of Cassava, the medium which contained the 2,4-D, BAP and IBA showed the highest callus induction rate, embryogenesis callus formation rate and somatic embryo formation in Cassava culture. In the medium with GA3 and myo-inositol, shoots were most vigorously regenerated from somatic embryos of Cassava. Our experiments confirmed that in vitro growth and multiplication of plantlets could depend on its reaction to the different medium composition, and this micropropagation techniques could be a useful system for healthy and vigorous plant production.

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Studies on Computer Optimization Techniques for Hydrophilic Vehicle Compositions

  • Lee, Chi-Ho;Shin, Young-Hee
    • Archives of Pharmacal Research
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    • v.11 no.3
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    • pp.185-196
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    • 1988
  • The inflence of hydrophilic vehicles on percutaneous absorption rate of griseofulvin was studied using intact skin of full thickness of hairless rat. The in vitro absorption rates were used as the characteristics for deciding the optimum formula of ointment vehicles. The optimum formula of vehicle compositions for maximum absorption rate was obtained from the polynomial regression equation and the two graphical techniques, contour graph and partial derivative graph. It was composed of sodium lauryl sulfate (1.65 W /W%), white petrolatum (16.5 W /W%), propylene glycol (12.0 W /W%), and stearyl alcohol (19.6W /W%). The experimental value obtained from the optimum formula and the prediction value were 33.99 and 33.87 ${\mu}g/\sqrt{min}$, respectively. From these results, it was believed that optimum formula for semisolid dosage forms could be obtained from the application of the optimization technique used in this study.

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