• Title/Summary/Keyword: in vitro maintenance

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Notch Is Not Involved in Physioxia-Mediated Stem Cell Maintenance in Midbrain Neural Stem Cells

  • Anne Herrmann;Anne K. Meyer;Lena Braunschweig;Lisa Wagenfuehr;Franz Markert;Deborah Kolitsch;Vladimir Vukicevic;Christiane Hartmann;Marlen Siebert;Monika Ehrhart-Bornstein;Andreas Hermann;Alexander Storch
    • International Journal of Stem Cells
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    • 제16권3호
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    • pp.293-303
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    • 2023
  • Background and Objectives: The physiological oxygen tension in fetal brains (~3%, physioxia) is beneficial for the maintenance of neural stem cells (NSCs). Sensitivity to oxygen varies between NSCs from different fetal brain regions, with midbrain NSCs showing selective susceptibility. Data on Hif-1𝛼/Notch regulatory interactions as well as our observations that Hif-1𝛼 and oxygen affect midbrain NSCs survival and proliferation prompted our investigations on involvement of Notch signalling in physioxia-dependent midbrain NSCs performance. Methods and Results: Here we found that physioxia (3% O2) compared to normoxia (21% O2) increased proliferation, maintained stemness by suppression of spontaneous differentiation and supported cell cycle progression. Microarray and qRT-PCR analyses identified significant changes of Notch related genes in midbrain NSCs after long-term (13 days), but not after short-term physioxia (48 hours). Consistently, inhibition of Notch signalling with DAPT increased, but its stimulation with Dll4 decreased spontaneous differentiation into neurons solely under normoxic but not under physioxic conditions. Conclusions: Notch signalling does not influence the fate decision of midbrain NSCs cultured in vitro in physioxia, where other factors like Hif-1𝛼 might be involved. Our findings on how physioxia effects in midbrain NSCs are transduced by alternative signalling might, at least in part, explain their selective susceptibility to oxygen.

The fibronectin concentration that optimally maintains porcine satellite cells

  • Jae Ho Han;Si Won Jang;Ye Rim Kim;Hoon Jang;Kwan Seob Shim;Hyun Woo Choi
    • Animal Bioscience
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    • 제36권12호
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    • pp.1889-1897
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    • 2023
  • Objective: 'Cultured meat' has been suggested as means of solving the problems associated with overpopulation and gas emissions. Satellite cells are a major component in the production of cultured meat; however, these cells cannot be maintained in vitro over long periods. Fibronectin is a glycoprotein that affects biological processes such as cell adhesion, differentiation, and migration. Unfortunately, the characteristics of porcine satellite cells grown in a long-term culture when exposed to fibronectin-coated dishes are unknown. The objective of this study was to investigate the appropriate concentration of fibronectin coated dishes for proliferation and maintenance of porcine satellite cells at long-term culture. Methods: In this study, we isolated the satellite cells and fibroblast cells with pre-plating method. We next analyzed the cell doubling time, cell cycle, and rate of expressed paired box 7 (Pax7) and myogenic differentiation 1 (MyoD1) in porcine satellite cells cultured with 20 ㎍/mL of fibronectin-, gelatin-, and non-coated dishes at early and late passage. We then analyzed the proliferation of porcine satellite cells with various concentrations of mixed gelatin/fibronectin. We next determined the optimal concentration of fibronectin that would encourage proliferation and maintenance of porcine satellite cells in a long-term culture. Results: Doubling time was lowest when 20 ㎍/mL of fibronectin was used (as tested during an early and late passage). Levels of expressed Pax7 and MyoD1, assessed using immunocytochemistry, were highest in cells grown using fibronectin-coated dishes. The proliferation of gelatin/fibronectin mixed coatings had no significant effect on porcine satellite cells. The concentration of 5 ㎍/mL fibronectin coated dishes showed the lowest doubling time and maintained expression of Pax7. Conclusion: Fibronectin with 5㎍/mL effectively maintains porcine satellite cells, a discovery that will be of interest to those developing the next generation of artificial meats.

The Evaluation of Nutritive Value of Three Tropical Browse Species for Sheep Using in Vitro and in Vivo Digestibility

  • Yahaya, M.S.;Kibon, A.;Aregheore, E.M.;Abdulrazak, S.A.;Takahashi, J.;Matsuoka, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제14권4호
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    • pp.496-500
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    • 2001
  • In vitro and in vivo experiments were carried out to evaluate the nutritive value of three tropical browse species as assessed by DM and CP digestibility, and NDF and ADF degradability with incubation times (T) of 6, 12, 24 and 48h. During the in vivo digestibility experiment three male castrated sheep (age 16 - 25 months) with a mean liveweight of $11.5{\pm}0.9kg$ were placed in individual metabolism stalls and were allocated to one of the three browse species in a $3{\times}3$ Latin square design. The browse species were all leguminous and consisted of: Acacia sieberina (A. sieberina), Ficus polita (F. polita), and Ficus sycomorus (F. sycomorus). The mean DM and CP contents of F. polita were higher than for A. sieberina and F. sycomorus (p<0.05). In contrast the NDF and ADF contents of F. sycomorus were higher compared to the other species examined (p<0.05). The in vitro DM and CP digestibility, and NDF and ADF degradability observed at different stages of incubation were higher in F. polita followed by A. sieberina and F. sycomorus. The DM and CP digestibility at 48 h incubation were 72.92, 74.84 and 53.52% and 77.38, 77.68 and 63.64% for A. seiberina, F. polita and F. sycomorus, respectively. This shows that F. polita contains more soluble materials which ruminant can benefit from and hence has more feeding value. The fermentation of F. sycomorus was slower for all the nutrients evaluated due to the presence of more fibre. Similarly, higher in vivo digestibility coefficient of DM, CP, NDF, ADF and hemicellulose were observed for F. polita reflecting its higher values of CP, ether extract (EE) and hemicellulose associated with lower values of NDF and ADF. Higher DMI and daily gain were recorded in sheep during feeding of F. polita compared to the other species evaluated. The digestibility of all the nutrients examined were higher in the in vivo than in the in vitro trial except for CP and DM. Sheep showed no visual signs of toxicity throughout the study periods. These results showed A. seiberina, F. polita and F. sycomorus can sustain sheep on a maintenance diet and could as well be used as a supplementary feed to low producing animals during the tropical dry season. Further research is needed to ascertain the viability of using these browse species on a long-term basis.

Efficient and Reliable in vitro Regeneration System for Rubus Species as the Basis of Genetic Engineering

  • Kalai Katalin;Meszaros Annamaria;Denes Ferenc;Zatyko Jozsef;Balazs Ervin
    • Journal of Plant Biotechnology
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    • 제7권4호
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    • pp.241-246
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    • 2005
  • Factors affecting regeneration of different Rubus varieties (blackberry, raspberry and their hybrid) were examined and a reliable regeneration system was established. Media for stock plant maintenance were tested; different explants and media were investigated to find the best circumstances for the regeneration. The effect of the commonly used antibiotics was studied to determine the most suitable one for selection of the transformants. We found that both MS and LS media supplemented by $20\;gL^{-1}$ sucrose are suitable for the stock plant maintenance. The optimal hormone content for the stock plants is $0.125\;mgL^{-1}$ 6-benzylaminopurine (BAP) with $0.01\;mgL^{-1}$ indole-3- butyric acid (IBA). The highest regeneration rate was observed on medium containing MS salts with B5 vitamins complemented with glucose, sucrose, maltose, $10\;gL^{-1}$ each, supplemented with benzylaminopurine riboside (BAR) ($2\;mgL^{-1}$) and indole-3-acetic acid (IAA) ($0.1\;mgL^{-1}$). The regenerated shoots appeared directly from the cut edges, without callus phase. Hygromycin and geneticin proved to be good selection agents for the Rubus explants, but due to their severe effect on the tissues we propose to use marker-free constructions for the transformation.

효소변형 전분기반 하이드로젤 모델 프리믹스 내 탑재된 커큐민의 소화과정 중 안정성 (Stability of the enzyme-modified starch-based hydrogel model premix with curcumin during in vitro digestion)

  • 강지현;노신정;이지영;김용노
    • 한국식품과학회지
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    • 제53권4호
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    • pp.365-374
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    • 2021
  • 본 연구에서는 커큐민 함유 에멀션의 안정성을 위해 필드하이드로젤로 쌀전분(RS), 1시간 효소처리한 전분(1GS), 24시간 효소처리한 전분(24GS), 96시간 효소처리한 전분(96GS)을 사용하였으며 이를 이용한 모델 프리믹스로 커큐민 탑재 필드하이드로젤 분말(FHP)을 제조하여 탑재된 커큐민의 안정성 및 in vitro 소화 후 커큐민 보유율의 변화를 관찰하였다. FHP는 커큐민 함유 에멀션분말(EMP)과 비교하여 재분산시 크리밍 현상없이 분산안전성을 보여주었다. 재분산 하이드로젤은 고온에서 녹으면서 오히려 액적(droplet)의 유착(coalescence)과 응집(flocculation)이 가속화되어 에멀션에 비해 열안정성이 떨어졌으나 UV 안정성은 RS-FHP와 1GS-FHP에서 유의적인 커큐민 보호 효과를 나타냈다. GS-FHP는 in vitro 소화 중 기름방울의 응집 및 유착이 발견되지 않았으며 소화 후 에멀션의 유상에 용해된 커큐민을 성공적으로 보호함을 확인하였다. 따라서, GS-FHP는 분말형태 프리믹스 개발에 새로운 소재로 사용 가능할 것이며 커큐민 외에 다른 다양한 소수성 기능성물질에 대해서도 적용을 확장할 수 있을것으로 생각된다.

Role of Gremlin-1 in Cancer

  • Park, Sin-Aye
    • 대한의생명과학회지
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    • 제24권4호
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    • pp.285-291
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    • 2018
  • Gremlin-1 (GREM1) has been defined as an antagonist of bone morphogenetic proteins (BMPs), particularly during embryonic development and tissue differentiation. However, recent studies have shown that GREM1 has BMPs-dependent or -independent functions in diverse human diseases. GREM1 plays a key role in the process of organ fibrosis, including lungs, kidneys, and so on. The GREM1-induced fibrosis typically promotes the development of other diseases, such as pulmonary hypertension, renal inflammation, and diabetic nephropathy. More recently, considerable evidence has been reported showing that GREM1 is involved in the promotion and/or progression of tumors in vitro and in vivo. It also performs an oncogenic role in the maintenance of cancer stem cells. Although GREM1 is known to function in a variety of diseases, here we focus on the role of GREM1 in cancer, and suggest GREM1 as a potential therapeutic target in certain types of cancer.

산지별 맥문동 부탄올분획물의 신경성장인자 유도 효과에 관한 비교 (Comparison of Nerve Growth Factor Induction by Butanol Fraction of Liriope platyphylla and Ophiopogon japonicus)

  • 강동호;김선여
    • 생약학회지
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    • 제39권2호
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    • pp.75-79
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    • 2008
  • Nerve growth factor (NGF) is a protein plays a major role in the development and maintenance of central and peripheral nervous system. Recent data suggest that reduced availability of NGF may play a significant role in the pathogenesis of diabetic $polyneuropathy.^{1)}$ In our previous study, steroidal saponin from Liriope platyphylla showed neurotrophic effect by stimulation of NGF synthesis and activation of tyrosine kinase $signaling.^{2)}$ In this study, we examined the neurotrophic effect of Liriope platyphylla (LP); which was from Mylyang(MYL) and Cheongyang(CHE), and Ophiopogon japonicus (CHI) on in vitro and in vivo model for the comparison of their NGF induction. We quantitatively analyzed spicatoside A in the LP and CHI by HPLC. And we investigated the correlation between the contents of spicatoside A and NGF induction efficacy on PC 12 cells and mouse serum. These results suggest that spicatoside A may enhance NGF induction in animal model.

Regulatory T Cell Therapy for Autoimmune Disease

  • Ha, Tai-You
    • IMMUNE NETWORK
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    • 제8권4호
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    • pp.107-123
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    • 2008
  • It has now been well documented in a variety of models that T regulatory T cells (Treg cells) play a pivotal role in the maintenance of self-tolerance, T cell homeostasis, tumor, allergy, autoimmunity, allograft transplantation and control of microbial infection. Recently, Treg cell are isolated and can be expanded in vitro and in vivo, and their role is the subject of intensive investigation, particularly on the possible Treg cell therapy for various immune-mediated diseases. A growing body of evidence has demonstrated that Treg cells can prevent or even cure a wide range of diseases, including tumor, allergic and autoimmune diseases, transplant rejection, graft-versus-host disease. Currently, a large body of data in the literature has been emerging and provided evidence that clear understanding of Treg cell work will present definite opportunities for successful Treg cell immunotherapy for the treatment of a broad spectrum of diseases. In this Review, I briefly discuss the biology of Treg cells, and summarize efforts to exploit Treg cell therapy for autoimmune diseases. This article also explores recent observations on pharmaceutical agents that abrogate or enhance the function of Treg cells for manipulation of Treg cells for therapeutic purpose.

Role of Unstable Phenanthrene-Degrading Pseudomonas species in Natural Attenuation of Phenanthrene-Contaminated Site

  • Prakash, Om;Lal, Rup
    • 한국미생물·생명공학회지
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    • 제41권1호
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    • pp.79-87
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    • 2013
  • An unstable yet efficient phenanthrene-degrading bacterium strain Ph-3 was isolated from a petroleum-contaminated site at the Mathura Oil Refinery, India. The strain was identified as Pseudomonas sp. using a polyphasic approach. An analysis of the intermediates and assays of the degradative enzymes from a crude extract of phenanthrene-grown cells showed a novel and previously unreported pattern of 1, 2-dihydroxy naphthalene and salicylic acid production. While strain Ph-3 lost its phenanthrene- degrading potential during successive transfers on a rich medium, it maintained this trait in oligotrophic soil conditions under the stress of the pollutant and degraded phenanthrene efficiently in soil microcosms. Although the maintenance and in vitro study of unstable phenotypes are difficult and such strains are often missed during isolation, purification, and screening, these bacteria constitute a substantial fraction of the microbial community at contaminated sites and play an important role in pollutant degradation during biostimulation or monitored natural attenuation.

The effect of mushroom components on the proliferation of HeLa cell line in vitro

  • Chung, Kyu-Sun
    • Archives of Pharmacal Research
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    • 제2권1호
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    • pp.25-33
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    • 1979
  • In order to find out nutritional effects of Korean edible mushrooms on the multiplication of tissue cells, the alcohol extracts and acid bydrolysates of eleven species of mushrooms were added to Earle's ESS. A comparison of the respective multiplications of HeLa cells in this solution and in control solution of TC=199. Yielded the following result: The alcohol extracts and acid hydrolysates of a Coprinus comatur, Agaricus campestris, Agaricus bisporus, Lentinus edodes, Tricholoma matsutake, Pleurotus ostretus, Ramaria botrytis and Pholiota nameko influenced favorably the maintenance of the normal form and monolayer of HeLa cells. The growth curves of HeLa cells in the cultures containing, respectively, the alcohol extracts and acid hydrolysates of these eight mushrooms showed that five species, i.e., Coprinus comatus, Agaricus compestris, Agaricus bisporus, Lentinus edodes and Tricholoma matsutake effected an excellent multiplication and that the other three species were less effective than those five species. As to the effects on the cell multiplication, no marked difference was observed between the alcohol extracts and the acid hydrolysates of the mushrooms tested.

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