• Title/Summary/Keyword: 분화

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Study on the simplifying antibody cocktail technique for isolation of human mesenchymal stromal cells (hMSCs) (사람 Mesenchymal stromal cell(hMSC) 분리를 위한 간소화된 방법에 대한 연구)

  • Park, Jung-Hyun;Kim, Kyoung-Hwa;Lee, Yong-Moo;Ku, Young;Rhyu, In-Chul;Han, Soo-Boo;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.34 no.1
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    • pp.93-100
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    • 2004
  • 많은 연구들에서 hMSC를 얻기 위해 centrifugation, fluoroscence activated cell sorter(FACS), magnetic activated cell sorter(MACS)가 이용되어져 왔다. 그러나 centrifugation만을 이용한 경우 순도가 떨어지며 FACS나 MACS의 경우에는 비용, 시간이 많이 드는 단점이 있다. 따라서 이 연구에서는 antibody cocktail을 이용하여 hMSC를 좀더 쉽게 얻어내는 방법에 대해 알아보았다. 사람의 골반에서 12G의 바늘을 이용하여 골수를 흡입한 후 heparin이 들어있는 시험관에 넣고 처리과정을 시행하기 전에 냉장고에 보관하며 가능한 한 빨리 처리 과정을 실시한다. 얻은 골수에 적당량의 RosetteSep( Stemcell Technologies)을 첨가한 후 실온에서 20분간 반응시킨다. 그 후 적당량의 Ficoll-paque위에 골수와 RosetteSep의 혼합물을 섞이지 않게 올리고 원심분리를 이용하여 원하는 세포층을 얻어낸다. 이 세포층을 따로 분리한 뒤 배양한다. 배양 시 세포가 80%이상 차기 전에 계속 passage를 시행하며 배양한다. 이는 세포가 밀도가 높아져 원치 않는 세포로 분화되는 것을 막기 위함이다. 배양된 세포가 다양한 분화능력을 가지고 있는지 알아보기 위해 세 가지로 분화를 유도하였다. 적절한 배지와 적절한 환경에서 배양함으로써 얻어진 세포를 osteoblast, chondroblast, adipocyte로 분화를 유도하였다. 분화된 세포가 원하는 형질의 세포로 분화되었는지를 확인하기 위하여 osteoblast의 경우 alizarin red staining, alkaline phosphatase activity, chondroblast의 경우 toluidine blue staining, adipocyte의 경우 Oil-Red-O staining으로 염색하여 분화를 확인하였다. 분리해낸 세포는 각각 세 가지 세포로 분화가 되었으며 이는 RosetteSep이 hMSC를 성공적으로 분리해냈다는 것을 보여준다. 그러나 모든 세포가 분화를 보이지는 않았으며 따라서 hMSC의 순도를 높이기 위한 연구가 더 필요하다. RosetteSep을 이용하면 다른 방법들 보다 쉽게 hMSC를 얻을 수 있으나 기존의 방법과 순도의 측면에서 더 비교할 필요가 있다.

Plant Regeneration from Unpollinated Ovary Culture in Allium tuberosum Rottl (부추의 미수분 자방배양에 의한 식물체 재분화)

  • 윤수진;손재근;권용삼
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.1
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    • pp.37-40
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    • 1999
  • This study was carried out to determine the optimal conditions for the production of plants derived from the unpollinated ovary culture of Chinese chive (Allium tuberosum Rottl.). The Chinese chive collected from Korea showed much higher frequency of plantlet formation than those from Japan in the culture of unpollinated ovaries. Among the collections, 'Youngiljaerae' showed the highest frequency of plantlet formation. The MS basal medium was superior to B/sub 5/ in plantlet formation. The ovaries inoculated on the 2,4-D-free medium were directly induced plantlets without callus formation. Floral parts inoculated as a unit played important roles in callus formation and plant production. The frequency of callus and plantlet formation was higher in the culture of ovary with anthers than that of ovary alone.

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Effect of Isomers of Conjugated Linoleic Acid on Porcine Preadipocyte Differentiation (Conjugated Linoleic Acid(CLA) 이성체가 돼지 지방전구세포의 분화에 미치는 영향)

  • Moon, H.S;Chung, C.S.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.967-974
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    • 2004
  • The current study was undertaken to determine the effect of various conjugated linoleic acid (CLA) isomers on differentiation of pig preadipocyte during culture. Preadipocyte(stroma-vascular cell) was isolated from the backfat of newborn pigs and cultured to differentiate into mature fat cell. Different doses of CLA isomers were treated to the culture media at different times. Cell differentiation was determined by measuring the glycerol3-phosphate dehydrogenase activity of the cultured preadipocytes. Twenty and fifty $\mu$M of trans110_cis 12 isomer of CLA inhibited differentiation of pig preadipocyte whereas cis9-cis II isomer stimulated the differentiation. Both cis9-transII and trans9-trans11 isomers showed no effect. Effect of CLA isomer was more evident at the early stage of culture(day 0-8), than the late stage(day 8-14). These results suggest that each CLA isomer has different effect on pig preadipocyte differentiation.

Associations between Youth Self-Differentiation, Perceived Parental Control, and Psychological/Behavioral Problems -Mediating effects of Self-differentiation- (청소년의 자아분화 수준, 부모통제에 대한 지각과 심리 및 행동문제의 관계 -자아분화의 매개효과를 중심으로-)

  • Chang, Kyung Moon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.2
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    • pp.845-854
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    • 2014
  • This study investigated the associations between youths' perceived parental control, psychological/behavioral problems, and self-differentiation in a sample of 355 middle and high school students. The sex differences in these variables, and the influences of perceived parental control and self-differentiation on youths' psychological and behavioral problems were analyzed. The mediating effects of self-differentiation between parental control and youths' psychological/behavioral problems, and between psychological and behavioral problems were also investigated. Some sex differences in the variables were found. Youths' psychological and behavioral problems were influenced by self-differentiation and parental psychological control. The mediating effects of self- differentiation were significant either complete or partial way.

The family differentiation level, quality of family relationship, and intimacy (가족분화수준, 가족관계의 질 및 친밀감간의 관계)

  • Nam, Soonhyeon;Han, Seongyeul
    • Korean Journal of Culture and Social Issue
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    • v.8 no.2
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    • pp.33-49
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    • 2002
  • The present study investigated the relationship among the level of family differentiation, quality of family relationship, and intimacy with their partners in 381 adult children. First, this study examined the relationship between the level of family differentiation and quality of family relationship. The result showed that the level of family differentiation was positively and significantly correlated to quality of family relationship. This finding indicated that family differentiation may be used for an objective and useful index to measure the dynamics of emotional system and the level of family-of-origin emotional health. Second, the influence of the family of origin was studied through exploring the relationship between the level of differentiation of family of origin, and adult children's intimacy with their partners. It was found that the family differentiation level was positively and significantly related to adult children's intimacy with their partners at each subsystem, but family differentiation levels of parents more influenced on adult male's intimacy than on adult female's intimacy with their partners.

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Factors Affecting Plant Regeneration from Seed-Derived Calli in Italian ryegrass (Lolium multiflorum Lam.) (이탈리안 라이그래스의 종자배양에 있어서 식물체 재분화에 미치는 몇 가지 요인)

  • 이효신;강경민;조진기
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.323-328
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    • 2001
  • In order to optimize the conditions of callus induction and plant regeneration in seed culture of Italian ryegrass, the effects of basic medium and carbon sources on seed culturability and genotypic difference of plant regenerability were investigated. MS medium was better than N6 and B5 medium in enhancing callus growth and plant regeneration. Sucrose was superior to maltose in plant regeneration as carbon source in the medium. The genotypic difference in plant regenerability was obvious among four cultivars of Italian ryegrass tested. 'Rio' and 'Jeanne' showed to have higher regenerability with the frequency of 38% and 56%, respectively.

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Plant Regeneration from the Stem Tissue of Orostachys japonicus A. Berger (바위솔의 줄기조직으로부터 식물체 재분화)

  • 최상욱;남상해;양기종;조무제;양민석
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.65-68
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    • 1994
  • Plant regeneration from the stem tissue of Orostachys japonicus A. Beiger was investigated. The calli derived from shoot apex when apex when cultured on Murashige and Skoog (MS) medium supplemented with 4mg/L 2,4-dichlorophenoxyacetic acid (2,4-D)and 2 mg/L benzyl aminopurine (BAP). The calli were developed into shoot to MS medium with 0.5mg/L NAA and 2mg/L and into root with 1mg/L kinetin. The reddish pigment which might be essential for the rootregeneration was observed in the tip of regenerated root.

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Effects of Polyamines on Adventitious Shoot Regeneration from Populus Leaf Segments (포플라 잎절편의 부정아 분화에 미치는 Polyamine의 영향)

  • 김성호
    • Journal of Plant Biology
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    • v.35 no.1
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    • pp.17-23
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    • 1992
  • Effects of polyamines and their biosynthetic inhibitors on adventitions shoot regeneration of Populus leaf segments were investigated. Polyamine inhibitors such as difluoromethyl arginine (DFMA), difluoromethyl ornithine (DFMO), and dicyclohexylamine (DCHA) decreased the fresh weight of cultured leaf segment and the rate of adventitious shoot regeneration. The inhibitory effects of DCHA were stronger than any other oplyamine inhibitors, and the rest were in the order of DFMO and DFMA. The inhibitory effects of these inhibitors were lessened or disappeared by the addition of polyamines, among which spermidine was the highest in its effect. Therefore, it is suggested that the spermidine may be related to the adventitious shoot regeneration of Populus leaf segments.gments.

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Comparison of Callus Formation Ratios from Seed Explants, Callus Sizes and Regeneration Efficiency Among Several Ochardgrass (Dactylis glomerata L.) Varieties (오차드그래스(Dactylis glomerata L.)의 품종에 따른 종자유래의 캘러스 형성률과, 캘러스 크기 및 식물체 재분화 효율의 비교)

  • 배은경;이인애;김기용;이병현;손대영;이효신;정민섭;조진기
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.2
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    • pp.93-100
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    • 2002
  • Comparison results of callus formation ratio from seed explants, callus sizes, regeneration ratios from callus and regeneration efficiency [calculated by following formular; callus formation ratio(%) ${\times}$ regeneration ratio(%)/100] for 27 ochardgrass (Dactylis glomerata L.)varieties imported and Hapsung 2 developed in Korea are as follows; 1. Among ochardgrass varieties showing more than 50% callus formation ratios, the descending order of callus formation ratio after bedding the seed explants for 4 weeks was 93M>Sparta> Pizza>Condor>Lidaglo>Glorus>Hapsung 2>Frode. 2. The callus sizes after bedding for 4 weeks were in the range of $\Phi$ 0.43cm~4.2cm in which there was 10 times size difference between the largest one and the smallest one but most of them were between ${\Phi}2.5cm~4cm$. 3. The regeneration ratio from callus among varieties were in the range of 0~36% and descending order of the upper 6 varieties was Plano>Akimidori>Justus>Lidacta>Currie>Hall mark. 4. The regeneration efficiency which is calculated by the ratios of regeneration from seed explant numbers was between 0 to 17.4% among which Justus showed the highest value in the 4-week treatment. 5. The correlation between callus formation ratios and the callus sizes, callus formation ratios and regeneration efficiency, and callus sizes and regeneration efficiency were r=0.5765, r=0.6365 and r=0.6246, respectively in 4-week callus and all the correlations were significant on the 1% level. 6. In 6-week callus, the descending order callus formation ratios from seed explants fur the best 6 varieties was Condor>Sparta>93 M>Justus>Potomac>Lidaglo>Frode. 7. The callus sizes formed were between ${\Phi}1.5~5.7cm$ in which Sparta, the largest one of ${\Phi}5.7cm$ was five times larger than the smallest one. The callus size of the control variety, Hapsung 2 was ${\Phi}3.8cm$, which belonged to a larger size. 8. Regeneration ratio showed a great deviation among varieties from 6-week old calli by showing from 0% to 100% in which all the calli were regenerated in Piano while no callus was regenerated in Juno. 9. The range of regeneration efficiency was between 0~28% among varieties in which the values from 6-week callus treatment were larger than those from 4-week callus treatment. Especially, the value of Potomac in 6-week was 3 times larger than that in 4-week. 10. The correlation between callus formation ratios and the callus sizes, callus formation ratios and regeneration efficiency, and callus sizes and regeneration efficiency were r=0.5369, r=0.6683 and r=0.5937. respectively in 6-week callus, and all the correlations were significant on the 1% level.

Effects of glutamine and AgNO3 on plant regeneration of Sedum sarmentosum (돌나물의 식물체 분화에 미치는 Glutamine과 AgNO3의 영향)

  • Ahn, Jeong-Ho;Kim, Hyun-Soon;Lee, Seung-Yeob
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.18-22
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    • 2009
  • This work was conducted to establish an efficient plant regeneration system for genetic transformation and the in vitro conservation of Sedum sarmentosum genetic resources. Effects of glutamine and $AgNO_3$ on plant regeneration between two genotypes were investigated using MS media supplemented with 0.2 mg/L NM and 3.0 mg/L BA. Calluses were formed on leaf explants placed on MS solid media supplemented with 3 mg/L 2,4-D and 1 mg/L BA. Calluses of Keumsan local strain produced shoots at a frequency of up to 100% after 50 days of culture on medium supplemented with glutamine. The highest number of shoots per callus was 17.6 at 350 mg/L glutamine. However, calluses of Wanju local strain gave rise to no shoots under the same culture conditions. Likewise, calluses of Keumsan local strain produced shoots at a frequency of up to 100% after 50 days of culture on medium supplemented with $AgNO_3$ whereas Wanju local strain sporadically produced shoots. The highest number of shoots per callus of Keumsan local stain was 16.1 at $15{\mu}M$ $AgNO_3$. Regenerated shoots were subcultured on hormone-free MS medium for rooting and shoot growth, and then 3-5 cm high plantlets were transplanted to the artificial soils comprising vermiculite and perlite, where they survived at a frequency of 88-100%. After being transplanted into upland soil:sand (1:1, v/v) in a greenhouse, regenerated plants showed a morphologically normal growth.