• Title/Summary/Keyword: in virto culture

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The Effect of Co-culture and Oxygen Concentration on In Virto Fertilization of Follicular Oocytes in Korean Native Cattle (공배양 및 산소농도가 한우 난포란의 체외발생에 미치는 영향)

  • 이재관;윤준진;황성수;윤종택;김창근;정영채
    • Korean Journal of Animal Reproduction
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    • v.22 no.1
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    • pp.43-50
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    • 1998
  • The effect of oxygen tension on embryonic development in co-culture was evaluated from the standpoint of the reduction of dissolved oxygen concentration by the oxygen consumption of feeder cells. Three co-culture systems using bovine oviductal epitherial cells (BOEC), African green monkey kidney cells (Vero cells) or buffalo rat liver cells (BRLC) have been compared in terms of development of bovine embryos derived from oocytes matured and fertilized in vitro. Among the co-cultured embryo, Vero cells su, pp.rted the highest developmental rate (29%) and the other two showed the similar rates. When the co-cultures were incubated in three different oxygen tension such as 5, 10, 20% oxygen atmosphere, embryos co-cultured with Vero cells at 10%-O2 resulted in the highest percentage of development. From the measurement of oxygen consumption of feeder cells, BRLC consumed 1.38 10-10 mg-O2/min/cell which was higher than 0.94 10-10 and 0.26 10-10mg-O2/min/cell for Vero cells and BOEC, respectively. Based on the oxygen consumption data, the phenomena of optimum oxygen tension required in embryo development in vitro has been analyzed, and we suggested that gas phase oxygen concentration, oxygen consumption rate of feeder cells and the number of feeder cells should be considered for the design of optimal co-culture system for effective fertilization of embryos in vitro.

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Plant Regeneration from Hypocotyl Explants of Several Species of Lycopersicon (토마토속 식물의 배축절편 배양에 의한 식물체 재분화)

  • 임학태;이건섭;용영록;송융남;김종화
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.137-143
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    • 1994
  • In an attempt to optimize the in vitro-regeneration conditions necessary for the genetic manipulation of tomato species, we examined several hybrid lines and wild species (peruvianum, pimpinellifolium, glandulosum) of Lycopersicon for. their, different regeneration ability. The basal medium used for callus growth and organogenesis was MSB (MS + B5) supplemented with three combinations of TDZ (Thidiazuron) 0.5mg/L+NAA 0.5mg/L, BA 2.0mg/L+NAA0.05 mg/L, and zeatin 3.0 mg/L + IAA 0.02 mg/L. In the genotype of Lycopenicon grandulosum, combination of TDZ and NAA was more effective in inducing shoot and root differentiation than those of BA and NAA or zeatin and IAA. When all genotypes tested were considered, however combination of zeatin and IAA was shown to be the best in shoot regeneration. Result indicate that callus and organogenesis of Lycopenicon species are dependent upon the hormone types and plant genotypes, but MSB medium with zeatin 3.0 mg/L + IAA 0.02 mg/L maybe appropriate for genotype-independent plant regeneration system of Lycopercicon species. We also tried TDZ as a cytokinin source in tomato tissue culture and found it highly significant in tomato regeneration system.

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Effects of Plant Growth substances on Organ Regeneration from in virto cultured Flower Buds of Mulberry(Morus alba L., Morus bombycis Koidz.) (뽕나무 화아의 기내배양에 있어서 생장조절물질이 기관분화에 미치는 영향)

  • Nam, Hyeok-U;Mun, Jae-Yu;Kim, Ho-Rak
    • Journal of Sericultural and Entomological Science
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    • v.30 no.1
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    • pp.1-7
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    • 1988
  • Flower buds of the mulberry(Morus alba L., Morus bombycis Koidz.) were cultured under different conditions such as basal media, and various concentrations of plant growth substances. Effects of the culture conditions on growth of the buds and organ regeneration were investigated and the result obtained are as follows: Murashige and Skoog(M.S.) medium was more effective on budding and growth of female(Keomseolppong) and male(Kaeryangppong) flower buds isolated directly from branches, compared to Greshoff & Doy(G.D.) and Wolter & Skog(W.S.) media. The growth of the female buds was promoted at higher concetration of benzyl amino purine(BAP) i.e., 2.0ppm. The female and male buds cultured after cuuting for seven days showed better growth than those without cutting treatment. The females and the males bloomed to form healthy stigmas and anthers, respectively, when cultured on M.S. media containing high Kinetin with low concentration of indole acetic acid(IAA).

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Effects of Dietary Lipid Sources and Meal Frequency on Growing Performance and Lipid Metabolism in Rats (식이지방의 종류와 식이급여형태가 흰쥐의 성장 및 지방대사에 미치는 영향)

  • 이재준
    • Journal of Nutrition and Health
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    • v.26 no.2
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    • pp.119-130
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    • 1993
  • This study was carried out to investigate the effects of dietary lipid sources and meal frequency on growing performance and lipid metabolism in Sprague-Dawley strain male rats. The experiment was conducted in 4$\times$2 factorial arrangement with 4 sources of dietary lipid(palm oil, beef tallow, soybean oil and hydrogenated soyben oil) and 2 meal frequencies(ad-libitum or meal feeding). During the 4-week feeding period the rats were fed either ad-libitum (AL) or a single daily 3-hour meal (09 : 02-12 : 00) during the dark period. In vitro cultures were carried out to study the cholesterol synthetic activity in hte liver prepared from rats used in feeding trials. And in vitro cultures were also carried out to study the lipogenic and lipolytic activity in the liver and adipose tissues prepared from rats used in feeding trials. Present data indicated that body weight gain, feed intake and FER of AS(ad-libitum+soybean oil)and AHS(ad-libitum+hydrogenated soybean oil) group were significantly(p<0.05) higher than those of the other groups. It was found that the feed intake of MF group was much less than that of AL group. Total body weight gained by MF group was only 60% of AL group. Growing performance was not affected by dietary lipid sources. The cholesterol synthetic activity in liver tissues culture was markedly(p<0.05) increased in MF diets, especially in soybean oil group. The lipogenic activity in liver tissues culture of MP(meal feeding+palm oil) and MHS(meal feeding+hydrogenated soybean oil)group was significantly (p<0.05) higher than that of AP(ad-libitum+palm oil) group and AHS(ad-libitum+hydrogenated soybean oil) group(p<0.05). Rats fed ad-libitum+beef tallow and fed meal feeding+beef tallow showed significantly(p<0.05) higher lipogenesis than the other groups. It was apparent that the lipogenic activity in liver tissues culture was not affected by dietary lipid sources and meal frequency. Lipolytic activity in liver tissue culture was significantly(p<0.001)different with meal frequency; MF group was higher than AL group, but was not greatly affected by dietary lipid sources. In the in vitro studies with adipose tissue, MF diets increased the lipogenic activity and inhibited the lipolytic activity in adipocytes. The lipogenic activity in adipocytes was significantly (p<0.001) different with dietary lipid sources and found to be beef tallow group was the higherst, but the sources of lipid in the diet did not exert any effect on the lipolytic acitivity.

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Chromosomal Analysis of Hanwoo Embryos by In Vitro Culture Condition (한우 체외 수정란의 체외 배양 조건에 따른 염색체 분석)

  • Choi, S.H.;Cho, S.R.;Han, M.H.;Kim, H.J.;Choe, C.Y.;Son, D.S.;Chung, Y.G.;Kim, S.K.;Sohn, S.H.
    • Journal of Embryo Transfer
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    • v.22 no.2
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    • pp.137-141
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    • 2007
  • Antioxidants were well known to be essential supplements in the complex media and serve as a reservoir of oxygen. In this study, Hanwoo COCs (cumulus oocytes complexes) were matured and developed in L-cysteine-TCM199 and analyzed metaphase chromosome. Maturation rate of Hanwoo COCs were 73.4%, 94.6% in 0.1% PVA, 0.1 mM L-cysteine, respectively and showed significantly different between the treatments (p<0.05). Blastocyst formation were revealed 20.3%, 10.0% in 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA, respectively. There were no significant difference among treatment groups. Metaphase chromosome were showed 18.3%, 12.0% in 5% FBS-TCM199, 0.1 mM L-cysteine, respectively and analyzable chromosome were 6.1%, 4.0% and had no differences between the treated groups. In the case of in vitro developmental stages, metaphase chromosome were showed 18.3%, 12.0% in $4{\sim}16$ cells stage, 43.1%, 13.0% in morulae stage and 94.8%, 100.0% in blastocyst stage. These results suggested L-cysteine has beneficial role for in virto maturation and development in Hanwoo COCs.

Regulation of hPTH Expression In Virto Using the Tetracycline Inducible Retrovirus Vector System (Tetracycline Inducible Retrovirus Vector System을 이용한 In Vitro에서의 인간 부갑상선 호르몬의 발현 조절)

  • Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.30 no.3
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    • pp.157-162
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    • 2006
  • Endogenous 84 amino acid parathyroid hormone(PTH) is synthesized as a pre-pro hormone by the chief cells of the parathyroid glands. Physiological actions of PTH include regulation of bone metabolism, renal tubular reabsorption of calcium and phosphate, and intestinal calcium absorption. In addition, PTH stimulates new bone formation by extraordinary stimulation of osteoblastic activity and decreasing calcium excretion by the kidney. In this study, we constructed and tested retrovirus vectors designed to express the human parathyroid hormone(hPTH) gene under the control of the tetracycline-inducible promoters. To increase the hPTH gene expression at turn-on state, woodchuck hepatitis virus posttranscriptional regulatory element(WPRE) sequence was also introduced into retrovirus vector at downstream region of either the hPTH gene or the sequence encoding reverse tetracycline-controlled transactivator(rtTA). Transformed primary culture cells(porcine fetal fibroblast, PFF, chicken embryonic fibroblast, CEF) were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the hPTH gene expression level using two step RT-PCR and ELISA Higher hPTH expression($3{\tims}10^4\;pg/ml,\;5.3{\times}10^4\;pg/ml$) and tighter expression control(up to 8 fold) were observed from the vector in which the WPRE sequence was placed at downstream of the hPTH gene. The resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

Studies on the Development of Acid Tolerant and Superior Nitrogen Fixation Symbionts for Pasture on Hilly Land -II. Selection of Acid Tolerant R. meliloti in virto and Inoculation Effect in Soils (야산(野山) 목초지용(牧草地用) 내산성(耐酸性) 우수(優秀) 질소고정균주(窒素固定菌株) 개발(開發) -II. 내산성(耐酸性) R. meliloti 의 기내선발(器內選拔) 및 토양(土壤) 접종효과(接種效果))

  • Kang, Ui-Gum;Choi, Ju-Hyeon;Cho, Kang-Jin;Jung, Yeun-Tae;Cho, Moo-Je
    • Korean Journal of Soil Science and Fertilizer
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    • v.22 no.1
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    • pp.72-77
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    • 1989
  • A study was conducted to obtain acid tolerant and superior Rhizobium meliloti strain for alfalfa on hilly acid soils. With host plant, eight isolates of R, meliloti selected in the vicinity of Milyang were evaluated for their ability to establish symbiotic effectiveness in acidified tube culture medium and vermiculite pot with different urea levels. Among isolates "YA03" was characterized for the ability to manifest to acid tolerance in three different soils of which pH were 5.0, 6.0, and 7.5. The results obtained were summarized as follows: 1. Of eight isolates "YA03" and "YA09" performed nodule in the tube medium of pH5.0, and in the symbiotic effectiveness YA03 was superior to others. 2. Alfalfa growth and $N_2-fixing$ activity by the inoculation of "YA03" isolate were better than others at the level of urea 1.25mM as well as nonurea. 3. Application of urea with inoculation of "YA03" islate to alfalfa was one of the effective factors for symbiotic effectiveness. 4. In infertile soil of pH5.0 inoculation of R. meliloti "YA03" to alfalfa caused the increase of shoot dry matter of 320% as compared to the control, and the total amount of yield was 131% as much yield in moderate fertile soil of pH7.5. Finally R. meliloti "YA03" isolate was selected as an acid tolerant strain.

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