• 제목/요약/키워드: tissue-culture

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야생 흰 제비꽃(Viola patrinii DC.) 배양세포의 생성과정 중 단백질 및 아미노산의 함량변화 (Alterations in Protein and Amino acid Contents During Growth on Culture Cells of Wild Viola (Viola patrinii DC.))

  • 정용모;임현희;조영수;정정한;이재헌;서정해;권오창
    • 식물조직배양학회지
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    • 제27권3호
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    • pp.181-184
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    • 2000
  • Friable callus와 compact callus를 배양 단계별로 단백질을 정량한 결과 friable callus가 가장 낮았고 배양 0주째부터 단백질 함량이 서서히 증가하여 배양 3주째 최대함량이 되었으며, 4주째부터는 감소하기 시작했다. 전기영동법으로 단백질의 패턴을 조사한 결과 compact callus와 friable callus간에 특징적인 차이를 보이는 단백질 밴드가 관찰되었다. Friable callus는 약 28 KD, 31 KD과 35 KD에서 특이적으로 나타났으며. compact callus에서는 30 KD부근의 밴드에서 많은 변화가 일어났다. 전체 단백질의 구성 아미노산은 배앙 2∼3주에 가장 높은 함량을 보였고, friable callus에서 그 함량이 가장 낮았다.

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소 체외수정란의 체외배양 및 이식후 생존성 (Viability of In Vitro Fertilized Bovine Embryos Following In Vitro Culture and Embryo Transfer)

  • 정희태;유재원;박연수;양부근;김정익
    • 한국수정란이식학회지
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    • 제9권3호
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    • pp.221-227
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    • 1994
  • This study was conducted to examine the condition of in vitro culture system and the viability after embryo transfer of in vitro matured-in vitro fertilized (IVM-IVF) bovine embryos. The in vitro development to the blastocyst stage was enhanced by supplying bovine serum albumin(BSA) to co-culture medium with bovine oviduct epithelial tissue(BOET) compared with that in medium supplemented with fetal bovine serum(FBS) (41.2% vs. 26. 3%, P<0.05). After transfer of IVM-IVF blastocysts into the uterine horn of recipient females (Aberdeen Angus), one was pregnant to term and produced a head of male Korean native calf. These results confirm that the in vitro development of IVM-IVF bovine embryos is affected with different protein source in co-culture with BOET, and IVM-IVF embryos can develop to term after in vitro culture and embryo transfer.

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Elimination of SPFMV from Virus-infected Sweet Potato Plants through Apical Meristem Culture

  • Kim, Young-Seon;Jeong, Jae-Hun;Park, Jong-Suk;Eun, Jong-Seon
    • Plant Resources
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    • 제7권3호
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    • pp.200-205
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    • 2004
  • Sweet potato infected with a viral disease (SPFMV) showed irregular chlorotic patterns, so called feathering associated with faint or distinct ring spots that have purple-pigmented borders. SPFMV was eliminated from sweet potato plants using meristem tip culture. MS medium supplemented with BAP (2mg/L) and NAA (0.05 mg/L) was used for shoot proliferation and 1/2 MS medium for rooting of the plants. Highest percentage of regenerated plants (60%) was obtained from the optimum size (0.3-0.5mm) meristem tips. Of these, 60% plants were found negative for SPFMV by RT-PCR. Virus detection by RT-PCR was found to be a reliable method. Meristem-tip culture to produce SPFMV-free quality sweet potato and virus detection by RT-PCR is an efficient, time saving and reliable method for production of SPFMV-free tissue culture raised plants.

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세포배양에 의한 고려인삼 성분의 생산 연구(II) (Production of Ginseng Saponins with Cell Culture(II))

  • 지형준;신국현;김현수;조희재
    • 생약학회지
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    • 제20권3호
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    • pp.162-169
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    • 1989
  • Tissue culture of the roots of Panax ginseng was carried out to enhance the production of ginseng callus as well as to increase its contents of ginsenosides. A long cylinder type callus mass was formed when cultured IK callus by rotary shaking culture method, the growth ratio of the callus being 7.71 which was approximately 4 fold higher than those obtained by other culture methods. Ginsenosides $Rg_1$, Re and $Rb_1$ could be detected from the callus mass by TLC, however, their total contents were revealed to be approximately 9% compared to that of the fresh ginseng root equivalent by HPLC analysis,.

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파벽(破壁) 영지포자가 고지방식이 섭취 마우스의 지방축적 및 체중감량에 미치는 영향 (Effect of Cell-Wall Broken Spores of Ganoderma lucidum(Leyss. ex.Fr.) Karst on the Lipid Accumulation and Body Weight Reduction in C57BL/6J Mice fed High-Fat Diet)

  • 박금주;강정일;김태석;이상윤;여익현
    • 생약학회지
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    • 제45권4호
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    • pp.346-353
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    • 2014
  • This study was performed to investigate the effects of cell-wall broken spores of Ganoderma lucidum on the lipid accumulation and body weight reduction in C57BL/6J mice. Six-week-old C57BL/6J mice were randomly divided into 5 groups and assigned to one of these groups; normal chew diet(Nor) group, high-fat diet(HFD) group, HFD plus spores of Ganoderma lucidum 800 mg/kg/day (HFD + GS/B) group, HFD plus cell-wall broken spores of Ganoderma lucidum 400 mg/kg/day (HFD + BGS/A) group and finally HFD plus cell-wall broken spores of Ganoderma lucidum 800mg/kg/day (HFD + BGS/B). The experimental groups which were treated oral co-administration with cell-wall broken(or original) spores of Ganoderma lucidum and HFD significantly attenuated accumulative body weight gain, compared with HFD group. Administration of these experimental materials also resulted in significant reduction not only the serum levels of total cholesterol, homocysteine but also the lipid accumulation in liver tissue. But in the almost of results the cell-wall broken spores of Ganoderma lucidum were evaluated superior than the original one. These results indicate that cell-wall broken spores of Ganoderma lucidum may inhibit the lipid accumulation in blood as well as liver tissue. Therefore it may be a valuable candidate for the therapy preventing obese induced hyperlipidemic symptoms.

Use of Neonatal Chondrocytes for Cartilage Tissue Engineering

  • KANG SUN WOONG;PARK JUNG HO;KIM BYUNG SOO
    • Journal of Microbiology and Biotechnology
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    • 제15권2호
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    • pp.259-264
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    • 2005
  • Transplantation of cultured chondrocytes can regenerate cartilage tissues in cartilage defects in humans. However, this method requires a long culture period to expand chondrocytes to a large number of cells for transplantation. In addition, chondrocytes may dedifferentiate during long-term culture. These problems can potentially be overcome by the use of undifferentiated or partially developed cartilage precursor cells derived from neonatal cartilage, which, unlike chondrocytes from adult cartilage, have the capacity for rapid in vitro cell expansion and may retain their differentiated phenotype during long-term culture. The purpose of this study was to compare the cell growth rate and phenotypic modulation during in vitro culture between adult chondrocytes and neonatal chondrocytes, and to demonstrate the feasibility of regenerating cartilage tissues in vivo by transplantation of neonatal chondrocytes expanded in vitro and seeded onto polymer scaffolds. When cultured in vitro, chondrocytes isolated from neonatal (immediately postpartum, 2 h of age) rats exhibited much higher growth rate than chondrocytes isolated from adult rats. After 5 days of culture, more neonatal chondrocytes were in the differentiated state than adult chondrocytes. Cultured neonatal chondrocytes were seeded onto biodegradable polymer scaffolds and transplanted into athymic mice's subcutaneous sites. Four weeks after implantation, neonatal chondrocyte-seeded scaffolds formed white cartilaginous tissues. Histological analysis of the implants with hematoxylin and eosin showed mature and well-formed cartilage. Alcian blue/ safranin-O staining and Masson's trichrome staining indicated the presence of highly sulfated glycosarninoglycans and collagen, respectively, both of which are the major extracellular matrices of cartilage. Immunohistochemical analysis showed that the collagen was mainly type II, the major collagen type in cartilage. These results showed that neonatal chondrocytes have potential to be a cell source for cartilage tissue engineering.

Production of tissue-type plasminogen activator from immobilized CHO cells introduced hypoxia response element

  • 배근원;김홍진;김기태;김익영
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2002년도 생물공학의 동향 (X)
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    • pp.257-260
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    • 2002
  • Dissolved oxygen level of cell culture media has a critical effect on cellular metabolism, which governs specific productivity of recombinant proteins and mammalian cell growth However, in the cores of cell aggregates or cell-immobilized beads, oxygen level frequently goes below a critical level. Mammalian cells have a number of genes induced in the lower level of oxygen, and the genes contain a common cis-acting element (-RCGTG-), hypoxia response element (HRE). By binding of hypoxia inducible factor-l (HIF-I) to the HRE, promoters of hypoxia inducible genes are activated, which is a survival mechanism. In this work, to develop a CHO cell capable of producing recombinant proteins in immobilization and high density cell culture efficiently, mammalian expression vectors containing human tissue-type plasminogen activator (t-PA) gene controlled by HRE were constructed and stably transfected into the CHO cells. In $Ba^{2+}$ -alginate immobilization culture, CHO/pCl/dhfr/2HRE-t-PA cells produced 2 folds higher recombinant t-PA activity than CHO/pCl/dhfrlt-PA cells without $CoCl_2$ treatment. Furthermore, in repeated fed batch culture, productivity of t-PA in immobilized CHO/pCI/dhfr/2HRE-t-PA cells was 121 ng/ml/day, total production of 0.968 mg/day at 11 days culture while CHO/pCIIdhfrlt-PA cells was 22.8 ng/ml/day. All these results indicate that HRE is very useful for the enhancement of protein productivity in mammalian cell cultures.

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Recent advances in seaweed seedling production: a review of eucheumatoids and other valuable seaweeds

  • Jiksing, Calvin;Ongkudon, McMarshall M.;Thien, Vun Yee;Rodrigues, Kenneth Francis;Yong, Wilson Thau Lym
    • ALGAE
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    • 제37권2호
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    • pp.105-121
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    • 2022
  • Modern seaweed farming relies heavily on seedlings from natural beds or vegetative cuttings from previous harvests. However, this farming method has some disadvantages, such as physiological variation in the seed stock and decreased genetic variability, which reduces the growth rate, carrageenan yield, and gel strength of the seaweeds. A new method of seedling production that is sustainable, scalable, and produces a large number of high-quality plantlets is needed to support the seaweed farming industry. Recent use of tissue culture and micropropagation techniques in eucheumatoid seaweed production has yielded promising results in increasing seed supply and growing uniform seedlings in large numbers in a shorter time. Several seaweed species have been successfully cultured and regenerated into new plantlets in laboratories using direct regeneration, callus culture, and protoplast culture. The use of biostimulants and plant growth regulators in culture media increases the seedling quality even further. Seedlings produced by micropropagation grew faster and had better biochemical properties than conventionally cultivated seedlings. Before being transferred to a land-based grow-out system or ocean nets for farming, tissue-cultured seedlings were recommended to undergo an acclimatization process to increase their survival rate. Regular monitoring is needed to prevent disease and pest infestations and grazing by herbivorous fish and turtles during the farming process. The current review discusses recent techniques for producing eucheumatoid and other valuable seaweed farming materials, emphasizing the efficiency of micropropagation and the transition from laboratory culture to cultivation in land-based or open-sea grow-out systems to elucidate optimal conditions for sustainable seaweed production.

기관배양을 통한 추간판 재생용 나노파이버 및 온도 감응성 지지체에 대한 검증 (Investigation of Nanofiber and Thermosensitive Scaffold for Intervertebral Disc through Organ Culture)

  • 이용재;신지원;신호준;김찬환;박기동;배진우;서형연;김영직;신정욱
    • 대한의용생체공학회:의공학회지
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    • 제28권4호
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    • pp.512-519
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    • 2007
  • The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.

조잭배양에 의한 씨마늘의 상업적 생산 (Commercial Production of Seed Garlic by Tissue Culture Technique)

  • 남상일;박주현;최종인;권기석;엄정식
    • 한국식물생명공학회:학술대회논문집
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    • 한국식물생명공학회 2002년도 춘계학술대회
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    • pp.33-40
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    • 2002
  • 동양물산 중앙기술연구소에서 개발한 다신초 (multiple shoot) 증식기술을 이용하면 1회 계대배양으로 신초의 수가 약 15배 증가하고 1년에 10회의 계대배양이 가능하므로 계산상 1개의 신초로부터 5,700억개 (1510)의 신초 생산ㅇ 가능하므로 유전적으로 동질의 씨마늘을 단기간에 대량증식시키는 것이 가능하다 이러한 기술을 우수한 형질을 가진 마늘종에 적용하여 현재 국내${\cdot}$외 여러 지역에서 적응성 및 안정성 생산성 검정 실험을 수행하여 산업화 진입을 준비하는 단계에 도달하였다. 수년에 걸친 검정 시험 결과 조직배양 씨마늘을 이용할 경우 최대 50% 이상의 증수효과는 물론 상품성이 증대되는 효과도 확인하여 우리 나라 마늘산업의 국제 경쟁력을 향상시키고 능민 소득 증대에 크게 기여할 수 있을 것으로 판단된다. 또한 조직배양을 통해 대량생산된 무병주 씨마늘을 농가 위탁, 계약재배 등을 통해 증식하여 실수요 농가에 보급하는 체계를 갖추게 된다면 타 주요작물에 비해 상대적으로 미진한 종구 보급체계도 확립될 수 있을 것이다. 더불어 국제경쟁력을 갖춘 기술력, 원가 등을 기반으로 국내뿐만 아니라 일본, 미국 등 국외지역에도 공급지역을 확대할 수 있을 것으로 기대된다.

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