• Title/Summary/Keyword: 계대배양

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Effect of Inorganic sale, Growth Regulators, and Thiamine on the Callus Growth and Plant Regeneration from long-term Cultured Solanum and Lycopenicon Genotypes (무기염류, 생장조절물질 및 타이아민이 장기간 계대배양한 Solanum과 Lycopnicon종의 캘러스 생장과 식물체분화에 미치는 영향)

  • Chang Yeon YU;Byong Ho CHANG;Dong Ha CHO
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.69-76
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    • 1994
  • Callus growth and shoot regeneration of Solanum and Lycopersicon depended on genotype, growth regulators, and thiamine concentration. L. peruvianum LA 1277 and L. peruvianum LA 1373 and PI 251301 had the greatest callus growth while L hirsutum LA 1777, L.esculentum 'Diego'and 'Red Plum' had the least callus growth. Lycopersicon penvianum genotypes were superior to L. esculentur genotypes in regenerating plants. MG medium was more effective in regenerating shoots than MS medium. A low level of IAA (0.2mg/L) and high level of BA (2 mg/L) resulted in the greatest shoot regeneration. Shoot growth varied depending on thiamine concentration and genotype. Shoot proliferation of Solanum ptycathum, Solanum nigrum, and L. peruvianum PI 199380 was best on medium with 20 mg/L thiamine. Regeneration of L. peruvianum PI 251301 and PI 128652 was better on medium with 30 and 10mg/L thiamine, respectively.

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소 체세포 핵이식에서 체세포 계대에 따른 염색체 이상

  • 최은주;이호준;김병정;민관식;윤종택
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.77-77
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    • 2002
  • 본 연구는 체세포 및 체세포 핵이식란의 염색체 이상에 체세포의 계대의 영향성을 조사하고자 하였다. 한우 성축의 귀 조직으로 얻어진 세포를 공여세포를 체외성숙 후 제핵된 난자에 핵이식을 실시하였으며, 1.9kv/cm, 20$\mu\textrm{s}$/2times의 전기자극으로 융합후 5$\mu\textrm{g}$/ml의 ionomycin에서 4min, 1.9mM 6-DMAP에서 4h동안 배양함으로써 활성화를 유도하였다. 핵이식란은 CRlaa에서 4일간 배양 후 8 -cell단계에서 중기상의 유도를 위하여 상기 배양액 1ml당 0.05$\mu\textrm{g}$ colcemid에서 6-8시간 더 배양하였다. 이후, 6% Fetal bovine serum이 함유된 1% sodium citrate용액에 20분간 저장처리 후, methanol 5 : aceticacid 1 : distilled water 4로 1차, methanol 3: aceticacid 1 로 조성된 2차, methanol 4 : acetic acid 3 : distilled water 1의 3차고정액으로 1분간 재 침지시켰다. 고정 처리가 완료된 slide는 4% Giemsa용액으로 염색한 후 광학현미경 하에서 핵형 양상을 검경하였다. 체세포의 5계대에서는 684개의 spreads를 검경한 결과 염색체 수는 72%가 정상으로 60개이었고, 24%가 60개 이하였으며 4%가 60개 이상을 보였다. 10계대도 5계대와 비슷하여 71%가 정상, 26%가 60개 이하, 3%가 60개 이상이었고, 15계대에서는 55%가 정상이었고, 30%가 60개이하, 15%가 60개 이상을 보였다. 10계대 까지는 mixoploid의 비율의 변화가 없었으나 15계대에서 현저하게 늘어남을 볼 수 있었다. 또한 체외수정란과 핵이식란의 비교에서는 체외수정란은 250개의 spreads를 검경한 결과 염색체 수는 95.6%가 정상으로 60개이었고, 2.0%가 60개 이하, 2.4%가 60개 이상이었으나, 핵이식란은 204개를 검경하여 88%가 정상이었고, 4.9%가 60개이하, 7.1%가 60개 이상을 보임으로써, 핵이식란이 체외수정란에 비하여 염색체 이상의 비율이 높았다. 따라서 계대에 따라 체세포의 염색체이상의 비율이 상대적으로 증가하고, 체세포 핵이식에 따른 염색체 이상이 생길 수 있음을 알 수 있었다. (이 논문은 농림부 연구비에 의해 수행되었음)

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Change in the RAPD Band of Strawberry Depending on 2,4-D and NAA Treatment and the Number of Subcultures In Vitro (기내배양에서 2,4-D 및 NAA처리와 계대배양회수에 따른 딸기의 RAPD Band 변화)

  • Shim, Jai-Sung;Chung, Hae-Joon;Min, Byung-Hoon
    • The Journal of Natural Sciences
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    • v.8 no.2
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    • pp.147-152
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    • 1996
  • The 2,4-D + BA combination in MS medium showed high regenerating ability and induced compact callus from leaf and petiole segments of three strawberry cultivars, 'Bokyo', 'Suhong', whereas the NAA + BA combination resulted in friable callus. Band of RAPD products obtained with primer 212 from the callus was different from the band of mother plant when callus induced from leaf segment of 'Suhong' cultivar was maintained in MS medium containing NAA or BA for 8 months. The RAPD bands obtained from mother plants of 'Bokyo' and 'Yeobong' were different from that of callus maintained in the presence of MS liquid medium containing 2,4-D(0.2 mg/$\ell$) subcultured every two weeks for 6 months(12 subcultures).

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Orchardgrass의 종자유래 캘러스로부터 부정배형성과 식물체 재분화

  • 이효신;이병현;원성혜;김기용;김미혜;정동민;조진기
    • Proceedings of the Korean Society of Grassland Science Conference
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    • 1999.06a
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    • pp.73.2-74
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    • 1999
  • Orchardgrass의 종자배양 유래의 캘러스를 현탁배양하여 현탁배양기간별 부정배형성정도와 식물체 재분화율 등에 대한 몇 가지 실험을 수행한 바, 2주 간격으로 4회계 대배양 하였을 때 계대배양 횟수가 증가됨에 따라 식물체 재분화율이 증가되었다. 종자배양에서 형성된 캘러스의 현탁배양에서 모양이 둥근세포와 그들의 세포괴는 배양 30일 후에 최대치를 나타내었고, 그 이후는 감소하였다.(중략)

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Colony Size Distributions according to in vitro Aging in Human Skin Fibroblasts (피부 섬유모세포 노화에 따른 세포집락 크기의 분포)

  • Kim, Jun-Sang;Kim, Jae-Sung;Cho, Moon-June;Park, Jeong-Kyu;Park, Tae-Hyun
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.158-165
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    • 1999
  • Purpose : To investigate the percentage of colonies wi1h16or more cells distribution of human skin fibroblast according to in vitro aging, and to evaluate the relationship between percentage of colonies with 10 or more cells and in vivo donor age in human skin fibroblast culture. Material and Method : C1, C2, C3a, and C3b human skin fibroblast samples from three breast cancer patients were used as subjects. The C1, C2, and C3a donor were 44, 54, and 55 years old, respectively. C3a and C3b cells were isolated from the same person. Single cell suspension of skin fibroblasts was prepared with primary explant technique. One hundred cells are plated into 100m1 tissue culture flask and cultured for two weeks. The colony size was defined as colonies with 16 or more cells. The cultured cell was stained with crystal violet, and number of cells in each colony was determined with stereo microscope at $\times$10 magnification. Passage number of C1, C2, C3a and C3b skin fibroblast were 12th, 17th, and 14th, respectively. Results : Percentage of colonies with 16 or more cells of skin fibroblast samples decreased with increasing in vitro passage number. In contrast, cumulative population doublings of skin fibroblast sample increased with increasing in vitro passage number. Percentage of colonies with 16 or more cells also decreased with increasing population doublings in human skin fibroblast culture. There was strong correlation with percentage of colonies with 16 or more cells and population doublings En C3a skin fibroblast sampie. At the same point of population doublings, the percentage of colonies with 16 or more cells of the young C1 donor was higher level than the old C3a donor. Conclusion : The population doublings increased with increasing in vitro passage number but percentage of colonies with 16 or more cells decreased. The results of this study imply that percentage of colonies with 16 or more cell is useful as a indicator of in vitro human skin fibroblast aging and may estimate the in vivo donor age.

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Relationship with Passage Time of Human Dental Pulp Stem Cells from Supernumerary Tooth by Classification (과잉치 분류에 따른 치수유래줄기세포 계대 배양 시간의 연관성)

  • Shin, Yeoseob;Kim, Jongbin;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.43 no.4
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    • pp.419-426
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    • 2016
  • For this research 20 supernumerary teeth impacted in the maxillary anterior have been extracted and pulp cells have been collected from them. From the collected pulp cells, total of 17 (10 males, 7 females) have been selected as subjects. From this research, the run-time of successive culture of the cell from tooth number pulp tissue was $2.91{\pm}0.29$ days. From the gathering of cells from the initial pulp tissue until gaining 80% confluency took $4.53{\pm}0.94$ which was the longest. The following successive cultures took $2.73{\pm}0.32$ days. Average runtime for female was $2.81{\pm}0.27$ days whereas male had average runtime of $2.98{\pm}0.29$ days. Average run-time for inversion was $2.94{\pm}0.30$ days and for normal location, $2.80{\pm}0.20$ days. Average runtime was $2.92{\pm}0.31$ days and other forms took $2.88{\pm}0.22$ days. In the future, follow up research would be needed to evaluate the efficiency of the cells collected from the initial passage and the latter passage as stem-cells and taking into consideration the less than 3 days'time for the subculture, it could be concluded that the research efficiency and fast cultivation would be sufficiently effective.

Gene Expression of Supernumerary Dental Pulp Related to the Subculture Speed: A Pilot Study (계대 배양 속도가 다른 과잉치 치수유래 줄기세포 간 유전자 발현 특성)

  • Lee, Yookyung;Kim, Jongsoo;Shin, Jisun;Kim, Jongbin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.46 no.2
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    • pp.219-225
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    • 2019
  • The purpose of this study was to investigate the odontoblast gene expression related to the subculture speed of supernumerary dental pulp stem cells (sDPSCs). The stem cell is undifferentiated cells which has the ability to differentiate into various cells. Specific stimulation or environment induces cell differentiation, and these differentiation leads to bone or muscle formation. 20 sDPSCs were obtained from 20 children under aseptic condition. During the culture through the 10th passage, the third passage cells which showed short subculture period and 10th passage cells which showed long subculture period were earned. Each cell was divided into differentiated group and non-differentiated group. Quantitative real-time polychain reaction (q-RT-PCR) was performed for each group. The genes related to odontoblast differentiation, Alkaline Phosphatase (ALP), Osteocalcin (OCN), Osteonectin (ONT), Dentin sialophosphoprotein (DSPP) and Dentin matrix acidic phosphoprotein 1 (DMP-1), were measured. Differentiated cells showed more gene expression levels. Undifferentiated cells showed higher gene expression level in 10th passages but differentiated cells showed higher gene expression level in 3rd passages. Cells that showed faster subculture period showed relatively lower gene expression level except for OCN and DSPP.

Flower Bud Induction and Flower Regeneration from Ovary Cultures of Allium fistulosum L. (파(Allium fistulosum L.)의 자방배양으로 부터 화아발생 및 꽃의 분화)

  • 김재훈;최용의;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.263-266
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    • 1998
  • Flowergenic callus was induced from the ovary surface of Allium fistulosum L. cultured on MS medium containing 0.5mg/L NAA and 0.5mg/L BA or 0.5mg/L kinetin. After 3-4 weeks of culture, the flower buds were developed from flowergenic callus. The continuous production of flowergenic callus was proliferated, when subcultured on the medium containing 0.5mg/L NAA and 0.5mg/L kinetin. However, frequency of flower bud formation from flowergenic callus was decreased as the subculture was repeated. Histological observation reveals that the developmental pattern of flower bud from flowergenic callus was closely similar to that of natural flowers.

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Development of In vitro Systems for Heat Tolerance Screening and Tuberization of True Potato Seeds (감자의 耐暑性 器內檢定 및 眞正種子의 器內塊莖形成體系 開發)

  • 김현준;김화영;유승렬;김병현;김정간
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.71-75
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    • 1997
  • For the establishment of early selection of heat-tolerant clones through in vitro tuberization of true potato seeds, different temperature treatments for in vitro tuberization were investigated. The ratios of tuberization at 2$0^{\circ}C$ on var. Superior and DTO-33 treated with 5 mg/L of BAP and 500 mg/L of CCC, were 85% and 92%, respectively. At 3$0^{\circ}C$, the ratio of tuberization on DTO-33 was 37%, which revealed strongly heat-tolerant clone. In culture system of in vitro tuberization, the number of tubers per flask at 2$0^{\circ}C$ on non-subculture incubation was more than that on subculture incubation. The condition of non-subculture and short-day treatment for 4 weeks was good for production as 10.6 tubers per flask, which was very similar to that of long-day treatment. On the other hand, tuber diameter on long-day treatment was greater as 11.2 mm than on short-day treatment. Therefore, in vitro tuberization from true potato seeds could be induced under the condition of long-day treatment at darkness.

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