• Title/Summary/Keyword: Tissue Culture

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Development of Biocompatible Vascular Graft -Endothelialization of Small Vascular Graft- (생체적합성 인조혈관의 개발 -혈관내피화 인조혈관-)

  • 김형묵;이윤신
    • Journal of Chest Surgery
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    • v.29 no.4
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    • pp.373-380
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    • 1996
  • Prevention of thromboembolism is the most important task in the development of bioconpatible small caliber artificial vascular graft. In normal vessels, vascular endothelial cells maintain homeosatsis by secreting numerous factors. The aim of this study is to develope a method which Improves biocompatibility of small caliver polyurethane graft using endothelial cell culture technique, and ev luate the efTectiveness of extracelluar matrix for endothelization which was produced by cultured fibroblast. Methods ; Multiporous polyurethane tube of 3 mm diameter, 0.3 mm thickness was manufactured for vascular graft. Three mongrel dogs were intubated and internal jugular veins removed. Extracelluar matrix produced by cultured flbrobast which was obtained from dog's internal jugular vein were coated to the polyurethane graft. Then, endothelial cells extracted from Jugular vein were cultured and fixed on the extracelluar matrix layer of vascular graft. Endothelial cell coated vascular grafts were implanted to the carotid arteries of experimental dogs as interposed autograft. Implanted grafts were removed after 3 and 6 weeks. As a control, PTFE graft was interposed on carotid artery. These experiments demonstrated that extracelluar matrix produced by fibroblast can afford a base for endothelial cell linings of polyurethane graft. Although thrombosis were developed on autografted en othelial cell coated graft, 33% opening was noticed, and showed less adhesion to adjacent tissue layer. These findings suggest that fiboblast produced extracelluar matrix which can be used for edothelial cell lining vascular graft, and by improving the cultured endothelial cell function, there will be a new modality for reducing thrombosis on small vascular graft.

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Studies on the Production of Watermelon and Cantaloupe Melonjuice (수박 및 참외 쥬스 제조에 대하여)

  • Shin, Dong-Hwa;Koo, Young-Jo;Kim, Choung-Ok;Min, Byong-Yong;Suh, Kee-Bong
    • Korean Journal of Food Science and Technology
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    • v.10 no.2
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    • pp.215-223
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    • 1978
  • In Korea, the annual production of watermelon and cantaloupe melon is around 110,000 to 170,000 Metric Tons, and as the fruit does not keep well, studies were conducted to determine the feasibility of preservation in the form of natural juice or lactic fermented juice. The results obtained in these studies are summarized as follows: (1) The average yield of juice obtained from watermelon was 56.2%, and from cantaloupe melon 65.8%, of the fresh weight. (2) The colloidal components of watermelon juice separated from the juice by sedimentation within 24 hours. Cantaloupe melon juice gave a stable colloidal dispersion. (3) No change in the colour of the juices was detected by sensory evaluation or instrumental methods after they were treated at $100^{\circ}C$ for 5 minutes. (4) The addition of canesugar to give a total solids content of 11/13 Brix gave juices which were preferred by most tasters. (5) Lactic fermentation of natural juices pasteurised at $90^{\circ}C$ for 5 minutes, and inoculated with a pure culture of lactic acid bacteria proceeded without interference from competing microorganisms. (6) Sensory evaluation of lactic fermented juices indicated that 60% of tasters found the juices as acceptable or better than commercial fruit nectars at present on the market. (7) Taste panels showed a preference for natural melon juices over the lactic fermented juices. (8) The peroxidase activity of cantaloupe melon juice was higher than that of watermelon juice, with juice extracted from the core of the fruit showing a higher activity than that from other portions of the tissue. (9) Two types of peroxidase, of differing heat stability were detected in both juices. The more heat stable peroxidase had a decimal reduction time of 40 minutes at $80^{\circ}C$ and a z value of $11^{\circ}C$.

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Etiology of Community-acquired Bacteremia in Healthy Children (건강한 소아에서 지역사회 획득 균혈증의 원인균)

  • Kim, Hyung Tae;Jang, Hyun Oh;Moon, Jin Soo;Nam, Seung Yeon;Kim, Dong Wook;Lee, Chong Guk;Cho, Chong Rae
    • Clinical and Experimental Pediatrics
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    • v.48 no.7
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    • pp.716-722
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    • 2005
  • Purpose : A full view of the spectrum of all bacterial diseases in healthy children is essential to the establishment of public health priorities. Accurate information on the relative importance of the various pathogens in terms of the age of the affected patients, the site of infection and the case fatality rate are valuable to the clinician in choosing antimicrobial treatments. Methods : Fifty-nine episodes of bacteremia were analysed. Data were collected at Ilsan Paik Hospital from January 2000 to December 2003. Analysis of each collected episode included isolating pathogen from blood culture, diagnosis, hospital course, isolating pathogens from other tissue sites, and studying results of antimicrobial sensitivity tests. Results : Fifty-nine cases of community-acquired bacteremia were reviewed. The most common pathogen was Staphylococcus aureus(11 cases, 18.6 percent), followed by Salmonella(10 cases, 16.9 percent), E. coli(7 cases, 11.9 percent), Streptococcus pneumoniae(five cases, 8.5 percent), Streptococcus viridans(5 cases 8.5 percent). The most common diagnosis was bacteremia without an indentified focus(61 percent), followed by meningitis(12 percent), bacteremia with enteritis(10.2 percent) and bacteremia with urinary tract infection(8.5 percent). Salmonella was still an important causative agent of bacteremia. The relative importance of Haemophilus influenza and Streptococcus pneumoniae was lower than in other studies. The most common organism responsible for bacteremia without an identified focus was Staphylococcus aureus. The case-fatality was 3.4 percent for all cases of bacteremia. Conclusion : We reviewed the etiology of community-acquired bacteremia. These data may be useful in the establishment of public health priorities and serve as a reference for selection of antibiotics in the empirical therapy of suspected invasive bacterial infection.

Regulatory Mechanism of Lipopolysaccharide(LPS)-Induced Interleukin-8 Gene Expression in Mononuclear Phagocytic Cells (단핵식세포에서 내독소에 의한 인터루킨-8 유전자 발현 조절기전에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.462-474
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    • 1994
  • Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.

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Plant Regeneration and Protein Analysis from Cadmium Resistant Callus of Tobacco (Nicotiana tabacum cv. BY4) (담배 (Nicotiana tabacum cv. BY4)카드뮴 저항성 캘러스로부터 식물체 재생과 단백질 분석)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.7-13
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    • 2001
  • Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

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A Study on the Growth Inhibition of Salmonella by Secretory Immunoglobulin A in Human and Bovine Milk (인유(人乳)와 우유(牛乳) 중에 함유(含有)되어 있는 Secretory Immunoglobulin A의 Salmonella속(屬) 성장억제(成長抑制)에 관(關)한 연구(硏究))

  • Bae, Heoung Churl;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.21 no.2
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    • pp.122-132
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    • 1994
  • Bacteriostatic activity of secretory immumoglobulin A (SIgA) in human and bovine colostrums on enterotoxigenic type Salmonella was tested in the tissue culture medium. SIgA was used in $0.1{\sim}5.0mg/m{\ell}$ concentration with or without the addition of egg lysozyme tested for theirs bacteriostatic activites. 1. Bovine SIgA rich fraction with a large amount of $IgG_1$-dimer could be prepared from bovine colostrum of Holstein cows by anion exchange chromatography using DEAE-Sephadex A-50 and gel filtration on Sephadex G-200 and Sepharose 6B. 2. Human SIgA appeared to be the most bacteriostatic effect for all varieties of Salmonella in a range of $0.5{\sim}1.0mg/m{\ell}$ Bovine SIgA showed a marked bacteriostatic effect increased by increasing the concentration. Bovine IgG had not show bacteriostatic effect against both enterotoxigenic type Salmonella. Egg lysozyme as well as bovine SIgA also showed a marked bacteriostatic effect increased by increasing the concentration. 3. When the growth inhibition of human SIgA was tested by adding egg lysozyme with time interval, egg lysozyme showed bacteriostatic effect as compared with control. But human SIgA and adding with lysozyme showed slight the bacteriostatic effect. 4. When the growth inhibition of bovine SIgA was tested by adding egg lysozyme with time interval, all treatment against S. enteritidis showed bacteriostatic effect as compared with control In the case of S. typhinwrium, egg lysozyme showed a marked bacteriostatic effect as compared with control.

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Ginsenoside R $b_2$ and Rc Formation and Inorganic Elements Uptake in Ginseng Hairy Roots Cultures (인삼모상근 배양에서 Ginsenoside R $b_2$ 및 Rc 생성과 무기이온 흡수)

  • 양덕조;윤길영;최규명;유승희
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.461-468
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    • 2000
  • For the enhancement of ginsenoside production in hairy roots cultures of Panax ginseng, the uptake rate of inorganic elements and ginsenoside contents were investigated by different concentrations of about phosphorus (P $O_{4}$$^{[-10]}$ ) and nitrogen (N $H_{4}$$^{+}$, N $O_{3}$$^{[-10]}$ ) sources. According to increased phosphorus and nitrogen sources, the uptake rate of $Mg^{2+}$ and F $e^{2+}$ in ginseng hairy roots were significantly increased. The uptake rate of F $e^{2+}$ in 5.15 mM N $H_{4}$$^{+}$ was higher at 47.5% than that in 20.6 mM, whereas that of C $u^{2+}$ in 10.3 mM were higher at 123.1% than that in 41.2 mM. These results indicated that phosphorus and nitrogen sources act not only elevated growth of hairy roots but also the uptake-enhancement of the irons and other ions. The optimum concentration of phosphorus and nitrogen sources for the contents of free sugars were different to kinds of free sugars. The optimum concentration of phosphorus and nitrogen sources for the ginsenoside formation in ginseng hairy roots cultures were highest at the most low concentration of all. The contents of ginsenoside-R $b_2$and -Rc in 0.31 mM P $O_{4}$$^{[-10]}$ were increased to 44.7% and 29.9% than that in 0.62 mM P $O_{4}$$^{[-10]}$ , respectively. The contents of ginsenoside-R $b_2$ and -Rc in 5.15 mM N $H_{4}$$^{+}$ were increased to 21.7% and 31.9% than that in 10.30 mM N $H_{4}$$^{+}$, respectively. The contents of ginsenoside-R $b_2$and -Rc in 4.7 mn N $O_{3}$$^{[-10]}$ were also increased to 17.6% and 25.5% than that in 9.4 mM N $O_{3}$$^{[-10]}$ , respectively. These results indicate that enhancement of the ginsenoside formation in ginseng hairy roots was feasible by new medium modulation of concentration of phosphorus and nitrogen sources.rogen sources.

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Effects of Dietary Beef Tallow, Soybean Oil and Squid Liver Oil on Growth and Body Composition of the Korean Rockfish Sebastes schlegeli, and Biochemical Changes with Starvation (사료 지질원으로 우지, 대두유 및 오징어 간유 첨가에 따른 조피볼락 Sebastes schlegeli의 성장 및 체성분 변화와 절식시 체내 대사)

  • LEE Sang-Min
    • Journal of Aquaculture
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    • v.7 no.1
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    • pp.63-76
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    • 1994
  • The effects of different dietary lipids on growth, body chemical composition, and nutrient partition of the Korean rockfish (43 g of initial mean weight) were studied. Fish were fed three experimental diets containing 1) $4.5\%$ squid liver oil, 2) $1\%$ squid liver oil and $3.5\%$ beef tallow, and 3) $1\%$ squid liver oil and $3.5\%$ soybean oil. Growth and feed efficiency were detarmined during a 19-week feeding period at water temperature of $15.5^{\circ}C$, and at the end of which body chemical compositions were analyzed. Remained fish were fasted for 45 days, and four fish were taken at 6, 12 and 24 hours, and 3, 15 and 45 days after the fasting to determine tissue compositions and hematological changes. Growth, feed efficiency, protein and lipid retention efficiency, hepatosomatic index(HSI), viscerosomatic index (VSI), and chemical composition of whole body and dorsal muscle were not affected by the different dietary lipids. Liver moisture content of fish fed squid liver oil diet was higher than that of fish fed beef tallow or soybean oil diet. Liver lipid content of fish fed beef tallow diet was higher than that of fish fed the other diets. Fatty acid composition of dorsal muscle and liver were affected by the different dietary lipids ; high levels of 20 : 5n-3 and 22 : 6n-3 from fish fed the diet containing squid liver oil, 18 : 1 from fish fed the diet containing beef tallow, and high 18 : 2n-6 and 18 : 3n-3 from fish fed the diet containing soybean oil were observed. Both HSI and VSI of fish fed three diets decreased with time after the begining of starvation. Liver glycogen did not change during the first 15 days of starvation and decreased thereafter, and that was not affected by the different dietary lipids. Lipid and protein contents in the dorsal muscle of fish decreased up to 15 days of starvation and remained unchanged thereafter, these were not different from each other. Glucose, free fatty acid, triglyceride and phospholipid concentrations in Fish serum were varied for the first 15 days of starvation, after that the concentrations of fish serum remained relatively stable in all the treatment groups with prolonged starvation. The results indicate that Korean rockfish can utilize fish oil, animal fat or vegetable oil equally as energy source when n-3HUF A is adquate.

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EFFECTS OF $INTERFERON-\gamma$ ON COLLAGEN AND FIBRONECTIN SYNTHESIS IN PRIMARY CULTURED PERIODONTAL LIGAMENT CELLS ($Interferon-\gamma$가 치주인대 세포의 Collagen 및 Fibronectin의 합성과 Alkaline Phosphatase 활성에 미치는 영향)

  • Kim, Gwang-Seok;Sung, Jae-Hyun;Choi, Je-Yong;Ryou, Hyun-Mo
    • The korean journal of orthodontics
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    • v.23 no.2 s.41
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    • pp.229-248
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    • 1993
  • [ $Interferon-\gamma$ ] has been suggested as a cytokine of connective tissue stabilizer. In addition, it has also been demonstrated that this cytokine inhibited bone remodeling activities of the bone derived cells. In order to illuminate the effects of this cytokine in orthodontic force induced bone remodeling, it was administered to primary cultured periodontal ligament cells which have been known to have some osteoblast like characteristics. $Interferon-\gamma$ slightly decreased $[^3H]thymidine$ incorporation rate without a significant change in the total cellular DNA content up to 1000 U/ml, which meant these doses were not cytotoxic to the cell. Total protein synthesis was not influenced by various concentration of interferon-y whether it was determined by the $[^3H]proline$ incorporation rate or by the Lowry smethod. The effect of $interferon-\gamma$ on the individual protein was, however, differential, ie, it increased $[^3H]proline$ incorporation into the noncollagenous protein marginally, while it decreased $[^3H]proline$ incorporation into the collagen, so that it caused dose-dependent suppression of the relative collagen synthesis. On the contrary, the fibronectin synthesis determined by the ELISA was increased by 1000 U/ml of $interferon-\gamma$. The differential effects of the interferon-y on the collagen and fibronectin synthesis exhibited not only their protein level but also the steady state mRNA level. $Interferon-\gamma$ decreased steady state level of ${\alpha}1(I)$ procollagen mRNA significantly, while showing no significant changes in the fibronectin mRNA level. In addition to this, it was also found that indomethacin did not affect on the $interferon-\gamma$ induced collagen decrease in this cell, which meant prostaglandins were not involed in the process of $interferon-\gamma$ induced collagen decrease. So it can be concluded that the incubation of periodontal ligament cells with 1000 U/ml of $interferon-\gamma$ for 24 hr showed differential effects on the type I collagen and fibronectin gene expression. The decrease in relative collagen synthesis in the protein level was related with decrease in the steady state level of mRNA, while the increase in the fibronectin synthesis in the protein level was not correlated with the mRNA level.

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Strategy for Bio-Diversity and Genetic Conservation of Forest Resources in Korea (생물종(生物種) 다양성(多樣性) 및 삼림유전자원(森林遺傳資源) 보존(保存) 전략(戰略))

  • Park, Young Goo
    • Journal of Korean Society of Forest Science
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    • v.83 no.2
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    • pp.191-204
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    • 1994
  • Due to its topographic complexities and various climatical condition, Korea exhibits diverse forest types. Dominant tree species in this zone are Quercus spp., Betula spp., Zelkova spp., Fraxinus spp., Pinus densiflora, Pinus koraiensis, and Pinus thunbergii ete. Genetic conservation in forest species in Korea there are three ways ; one is in situ, other is ex situ and third is in-facility conservation. In situ conservation include that are the present status of conservation of rare and endangered flora and ecosystem, the reserved forest, the national and provincial park, and the gene pool of natural forests. Ex situ conservation means to be established the new forest from in situ forest stands, progeny and provenance test populations, seed orchard and clone banks, and gene conservation in-facility. As a tool for low temperature storage, several aspects on in vitro system were studied ; (1) establishment of in vitro cultures from juvenile and/or rejuvenated tissues, (2) induction of multiple shoots from the individual micropropagules, (3) elongation of the proliferated shoots. Studies on cold storage for short-and long-term maintenance of in vitro cultures under $4^{\circ}C$ in the refrigerator were conducted. For the cryopreservation at $-196^{\circ}C$, various factors affecting survivability of the plant materials are being examined. The necessity of gene conservation of forest trees is enlarged not only to increase the adaptability for various environments but also to gain the breeding materials in the future. For effective gene conservation of forest trees, I would like to suggest followings ; 1. Forest stands reserved for other than the gene conservation purposes such as national parks should be investigated by botanical and gene-ecological studies for selecting bio-diversity and gene conservation stands. 2. Reserved forest for gene pool should be extented both economically important tree spp. and non-economical species. 3. Reserved forest for progeny test and clone bank should be systematically investigated for the use of Ex situ forest gene conservation. 4. We have to find out a new methodology of genetic analysis determining the proper and effective size of subpopulation for in situ gene conservation. 5. We should develop a new tree breeding systems for successful gene conservation and utilization of the genetic resources. 6. New method of in-facility gene conservation using advanced genetic engineering should be developed to save time and economic resources. 7. For the conservation of species with short-life span of seed or shortage of knowledge of seed physiology, tissue culture techniques will be played a great role for gene conservation of those species. 8. It is are very useful conservation not only of genes but of genotypes which were selected already by breeding program. 9. Institutional and administrative arrangements including legistlation must be necessarily taken for gene conservation of forest trees. 10. It is national problems for conservation of forest resources which have been rapidly destroyed because of degenerating environmental condition and of inexperienced management system of bio-diversity and gene conservation. 11. In order to international cooperation for exchanging data of bio-diversity and gene conservation, we should connect to international net works as soon as possible.

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