• 제목/요약/키워드: in vitro clonal proliferation

검색결과 10건 처리시간 0.031초

Shoot Proliferation of Populus euramericana(Populus deltoides X P. nigra) through in vitro Tissue Culture

  • Kang, Ho-Duck;Moon, Heung-Kyu
    • Plant Resources
    • /
    • 제4권3호
    • /
    • pp.111-120
    • /
    • 2001
  • The efficiency of in vitro regeneration of four clones of Populus euramericana, Canada blanc, Eugenii, I-45/51, and Wisconsin #5, was examined. Cytokinins and the combinations with auxins affected the rate of regeneration from the explants of root segments, stem internodes, and leaf discs. Overall, BA and the combination with auxins were effective in root segments and leaf discs of the Canada blanc clone, whereas zeatin and the combination with auxins were important in stem internodes of the Wisconsin #5 clone. The highest number of shoots averaging 17.6 $\pm$ 0.47 from root segments in the Canada blanc clone,18.2 $\pm$ 3.0 from stem internodes in the Wisconsin #5 clone, and 17.8 $\pm$ 1.92 from leaf discs in the Canada blanc clone were obtained with 2.0 mg/1 BA, 2.0 mg/l zeatin combined with 0.2 mg/l IAA, and 0.5 mg/l BA combined with 0.05 mg/l 2,4-D, respectively. In particular, the addition of 2,4-D into cytokinin medium promoted shoot proliferation.

  • PDF

제브라피쉬(Danio rerio) 배아로부터 동형세포주 확립 (Establishment and Characterization of Clonal Cell Lines from Zebrafish, Danio rerio)

  • 이기영
    • 한국어류학회지
    • /
    • 제20권1호
    • /
    • pp.1-6
    • /
    • 2008
  • 제브라피쉬 배아로부터 확보한 세포주로부터 외부형태와 세포 크기에 따라 3종류의 동형세포주를 확립하였다. 활발하게 증식하는 안정된 세포주 및 이들로부터 확립된 동형세포주의 세포특성은 변하지 않고 지속적으로 유지되었으며, 안정된 세포주로부터 총 18개의 콜로니를 확보하여 배양한 다음 3종류의 동형세포주를 선별하여 세포 특성을 분석하였다. 대부분의 동형세포주는 약 80% 정도 정상적인 염색체(2N=50)를 가지고 있었으며 FACs 분석과 일치하였다. 배아로부터 확립된 동형세포주에 항체테스트 결과, vimentin에서 양성을 보이는 결과로 볼 때 확립된 세포주는 분화된 섬유세포임이 확인되었다. 이러한 결과는, 확립된 동형세포주를 이용한 유전자조작과 어류복제에의 활용도를 높일 수 있음을 시사한다.

Mesenchymal Stem Cell Lines Isolated by Different Isolation Methods Show Variations in the Regulation of Graft-versus-host Disease

  • Yoo, Hyun Seung;Yi, TacGhee;Cho, Yun Kyoung;Kim, Woo Cheol;Song, Sun U.;Jeon, Myung-Shin
    • IMMUNE NETWORK
    • /
    • 제13권4호
    • /
    • pp.133-140
    • /
    • 2013
  • Since the discovery of the immunomodulation property of mesenchymal stem cells (MSCs) about a decade ago, it has been extensively investigated whether MSCs can be used for the treatment of immune-related diseases, such as graft versus-host disease (GvHD). However, how to evaluate the efficacy of human MSCs for the clinical trial is still unclear. We used an MHC-mismatched model of GvHD (B6 into BALB/c). Surprisingly, the administration of the human MSCs (hMSCs) could reduce the GvHD-related mortality of the mouse recipients and xenogeneically inhibit mouse T-cell proliferation and $IFN-{\gamma}$ production in vitro. We recently established a new protocol for the isolation of a homogeneous population of MSCs called subfractionation culturing methods (SCM), and established a library of clonal MSC lines. Therefore, we also investigated whether MSCs isolated by the conventional gradient centrifugation method (GCM) and SCM show different efficacy in vivo. Intriguingly, clonal hMSCs (hcMSCs) isolated by SCM showed better efficacy than hMSCs isolated by GCM. Based on these results, the MHC-mismatched model of GvHD may be useful for evaluating the efficacy of human MSCs before the clinical trial. The results of this study suggest that different MSC lines may show different efficacy in vivo and in vitro.

귀에 존재하는 내인성 성체줄기세포 (Endogenous Stem Cells in the Ear)

  • 박경호
    • Korean Journal of Otorhinolaryngology-Head and Neck Surgery
    • /
    • 제56권12호
    • /
    • pp.749-753
    • /
    • 2013
  • Basically stem cells have characteristics of multi-potency, differentiation into multiple tissue types, and self-renew through proliferation. Recent advances in stem cell biology can make identifying the stem-cell like cells in various mammalian tissues. Stem cells in various tissues can restore damaged tissue. Stem cells from the adult nervous system proliferate to form clonal floating colonies called spheres in vitro, and recent studies have demonstrated sphere formation by cells in the tympanic membrane, vestibular system, spiral ganglion, and partly in the organ of Corti. The presence of stem cells in the ear raises the possibilities for the regeneration of the tympanic membrane & inner ear hair cells & neurons. But the gradual loss of stem cells postnatally in the organ of Corti may correlate with the loss of regenerative capacity and limited hearing restoration. Future strategies using endogenous stem cells in the ear can be the another treatment modality for the patients with intractable inner ear diseases.

An Efficient Plant Regeneration System for Sorghum bicolor - a Valuable Major Cereal Crop

  • Baskaran P.;Jayabalan N.
    • Journal of Plant Biotechnology
    • /
    • 제7권4호
    • /
    • pp.247-257
    • /
    • 2005
  • An efficient, rapid and large-scale in vitro clonal propagation of agronomically important Indian cereal crop genotypes (NSH27 & K5) of Sorghum bicolor (L.) Moench. by enhanced shoot proliferation in shoot tip segments was designed. MS medium fortified with plant growth regulators and coconut water markedly influenced in vitro propagation of Sorghum bicolor. In vitro plantlet production system has been investigated on Murashige and Skoog (MS) medium with the synergistic combination of 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), 5% coconut water and 3% sucrose which promoted the maximum number of shoots as well as beneficial shoot length. Subculturing of shoot tip segments on a similar medium enabled continuous production of more than 100 healthy shoots with similar frequency. When the healthy shoot clumps were cultured on MS medium fortified with 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), ${\alpha}$-naphthaleneacetic acid ($2.7\;{\mu}M$), ascorbic acid ($30.0\;{\mu}M$) and 5% coconut water, a rapid production of axillary and adventitious buds was developed after 8 wk culture. More than 300 shoots were produced 10 wk after culture. Rooting was highest (100%) on half strength MS medium containing 22.8 mM IAA. Micropropagated plants established in garden soil, farmyard soil and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics. These plants grew normally without showing any traits.

In Vitro Propagation Through Nodal Explants in Helicteres isora L., a Medicinally Important Plant

  • Shriram, Varsha;Kumar, Vinay;Shitole, M.G.
    • Journal of Plant Biotechnology
    • /
    • 제34권3호
    • /
    • pp.189-195
    • /
    • 2007
  • Helicteres isora is medicinally important plant effective against asthma, diabetes, hypolipidemia, HIV, besides a good source of diosgenin. Seed dormancy and low rate of natural fruit production make this plant a perfect candidate for developing an in vitro method useful for its clonal propagation and further biotechnological developments. This is the first report on in vitro production of this plant. Nodal explants obtained from aseptically germinated seedlings were cultured on MS medium (Murashige and Skoog 1962) fortified with indole-3-acetic acid (IAA) ($0.57-22.83\;{\mu}M$), indole-3-butyric acid (IBA) ($0.41-16.58\;{\mu}M$), 6-benzylaminopurine (BA) ($0.44-17.75\;{\mu}M$) and kinetin (Kin) ($0.46-13.94\;{\mu}M$) either singly or in combinations of IAA + BA, IAA + Kin and BA + Kin. Combinations of cytokinins (BA and Kin) were most suitable for multiple shoot induction and $13.94\;{\mu}M\;Kin\;+\;13.31\;{\mu}M\;BA$ was optimum (79% frequency) associated with high number of microshoots (7.1 shoots per explant) after 20 days of culture. Maximum shoot elongation and proliferation (10 shoots per explant with 4.8 cm average height) was achieved on MS media containing $2.32\;{\mu}M\;Kin\;+\;2.22\;{\mu}M\;BA\;+\;2.85\;{\mu}M\;IAA$. High rooting frequency (70%) was achieved on MS medium (1/2 basal strength) fortified with $4.14\;{\mu}M$ IBA, while activated charcoal showed inhibitory effects on rooting. Hardening was done with 76% survival rate and these plants were growing without any visual defects and morphologically mimicking the naturally growing plants.

In vitro Callus formation and Plant Regeneration of Epimedium koreanum Nakai

  • Yu, Chang-Yeon;Kim, Na-Young;Kang, Won-Hee;Kim, Myong-Jo;Heo, Kwon;Choi, Won-Cheol;Lee, Hyeon-Yong
    • 한국약용작물학회지
    • /
    • 제10권4호
    • /
    • pp.259-262
    • /
    • 2002
  • The efficiency of regeneration of callus and explants from leaf and stem disks of Epimedium koreanum was examined on the MS media containing 2,4-D, NAA, Kinetin, BA and TDZ. Calli were formed on the 2mg/l 2,4-D media at the rate of 32% from leaf discs and 52% from stems. No callus was produced on the media which are containing BA or TDZ alone. The combination of 2,4-D and BA showed the effect on the formation of callus. The combination of 2mg/l 2,4-D and 0.lmg/l BA in the MS media had produced the highest percentage of callus formation, 50% from leaf discs and 40% from stems, respectively. The combination of 2mg/l 2,4-D and 1mg/1 BA in the MS media had affected the formation of callus in the rate of 40% from leaf discs and 25% from stems. The combined plant growth regulators of 2,4-D and BA increased the formation of calli from leaf discs, but single treatment of 2,4-D showed the highest callus formation from stems. Multiple shoots from leaf discs were formed on the media containing NAA, BA, kinetin, and TDZ. The highest number of multiple shoots were obtained 0.1mg/l NAA combined with 1mg/l kinetin. As a result, leaf discs or stems can be used for the mass propagation of Epimedium koreanum, but stem elongation of shoots from calli was not easy.

고구마의 액아배양에서 생장조절물질이 기관분화 및 생장에 미치는 영향 (Effect of Growth Regulators on the Organ Differentiation and the Growth from the Axillary Bud of Sweetpotatoes in Vitro Culture)

  • 장병호
    • 한국작물학회지
    • /
    • 제29권4호
    • /
    • pp.401-408
    • /
    • 1984
  • 우리나라에서 고구마 장려품종인 홍미와 신미를 액아배양시 explant의 크기 NAA, BA 및 kinetin의 농도가 기관분화와 생육에 미치는 영향을 구명하고저 본 실험을 실시하였으며 실험결과를 요약하면 다음과 같다. 1. 5mm되는 explant 배양이 2mm explant 배양보다 기관분화 및 생육이 좋았으며 홍미는 MS 배양에 NAA 0.1mg/$\ell$와 kinetin 1mg/$\ell$을 조합처리할 때 신미는 MS배지에 NAA 0.1mg/$\ell$와 kinetin 1mg/$\ell$을 조합처리할 때와 kinetin 1mg/$\ell$, BA 0.1mg/$\ell$을 단독처리할 때 shoot의 분화가 좋았다. 2. 뿌리분화는 홍미와 신미가 비슷한 결과를 보였으며 NAA 0.lmg/$\ell$와 kinetin 1mg/$\ell$을 조합처이한 배지에서 뿌리수와 뿌리길이가 증가하였다. 3. BA의 농도가 증가함에 따라 비정상 식물체의 출현이 많았으며 한 개의 explant에서 여러개의 shoot의 길이가 짧았다. 4. 이차계대배양에서 shoot의 분화 및 생장은 NAA 0.1mg/$\ell$와 kinetin 1mg/$\ell$가 첨가된 배지에서 좋은 결과를 보였고 뿌리의 분화는 0.1 mg/$\ell$ BA에 첨가된 배지에서 가장 효과적이었다.

  • PDF

개방형 액체대량배양 시스템을 통한 유칼리나무 펠리타 선발목의 대량증식 (Application of Open-type Liquid Culture for Large-scale Production of Mature Plus Tree of Eucalyptus pellita)

  • 박소영;문홍규;김용욱;김선자;이재선
    • 한국산림과학회지
    • /
    • 제97권6호
    • /
    • pp.650-655
    • /
    • 2008
  • 유칼리나무 펠리타(Eucalyptus pellita) 선발목(5년생)의 기내 대량증식을 위해 1L 규모로 1) 고체배양(대조구), 2) 액체정치배양, 3) 개방형 액체배양 등 3가지 배양법이 시도 되었고, 그 결과를 토대로 배양규모를 10L로 확대하여 대량배양 가능성을 조사하였다. 1L 배양용기에서 4주간 배양한 결과, 공기가 공급 된 개방형 액체배양에서 가장 좋은 생장 결과를 얻었다. 이는 액체배지에 의한 원활한 염류 공급과 배양용기내 공기의 환기에 의해 얻어진 결과로 생각된다. 배양 규모를 10L로 확대하여 액체대량배양을 실시한 결과, 식물체의 생장은 대조구인 고체배양에 비교하여 마디 수에 있어서 370%, 엽 면적 3.6배, 그리고 신초 길이 3.3배 신장하는 결과를 보였다. 이는 개방형 액체대량배양 시스템이 유칼리나무 클론증식시 생산성을 향상시키는데 적합한 시스템임을 보여주는 결과라고 하겠다.

간헐적 침지 방식의 생물반응기 시스템을 이용한 유칼리 선발목 클론 대량증식 (Use of the Temporary Immersion Bioreactor System for Mass Production of Eucalyptus pellita Plus Tree)

  • 김선자;박소영;문흥규;이위영
    • 한국산림과학회지
    • /
    • 제99권1호
    • /
    • pp.125-130
    • /
    • 2010
  • 식물대량증식에서 생물반응기의 이용은 규모를 대량화하고 자동화 할수 있다는 점에서 생산비를 절감할 수 있는 방법 중 하나이다. 본 연구에서는 유칼리 펠리타 선발목의 대량증식 체계를 확립하기 위해 4가지의 생물반응기 배양시스템에서 유칼리의 생장을 비교하였다. 배양기내에 지지물(net)을 설치하고 매 4시간마다 30분씩 액체배지를 공급한 TIN 배양(Temporary immersion with net)에서 식물체 생장이 가장 좋았다. TIN 시스템하에서 자란 식물체는 동일한 방식에 net가 없는 TIX 배양(Temporary immersion without net)에서 자란 식물체와 비교하여 초장이 3배 이상 증가하였다. 게다가 TIN 시스템에서 생산 된 식물체는 총 엽록소 함량, 엽록소 a/b, 그리고 건물중 등도 증가하였다. 위와 같은 결과는 기내 유칼리나무 생장에 중요한 요인이 식물체가 적당한 간격으로, 그리고 적당한 시간동안 배지에 노출되어야하고, net의 이용이 필수적임을 보여준다. TIN 시스템은 유칼리 클론묘의 대량생산을 위해 최적의 시스템으로 산업화를 위한 유칼리나무 대량생산시 유용하게 이용될 수 있을 것으로 생각된다.