scholarly journals In Vitro Tissue Culture in Brachypodium: Applications and Challenges

2020 ◽  
Vol 21 (3) ◽  
pp. 1037 ◽  
Author(s):  
Alexander Betekhtin ◽  
Karolina Hus ◽  
Magdalena Rojek-Jelonek ◽  
Ewa Kurczynska ◽  
Candida Nibau ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Transformation Efficiency ◽  
Brachypodium Distachyon ◽  
Low Frequency ◽  
Transformation Techniques ◽  
Bioenergy Grasses ◽  
Embryogenic Potential ◽  
Gradual Loss ◽  
Made In

Brachypodium distachyon has become an excellent model for plant breeding and bioenergy grasses that permits many fundamental questions in grass biology to be addressed. One of the constraints to performing research in many grasses has been the difficulty with which they can be genetically transformed and the generally low frequency of such transformations. In this review, we discuss the contribution that transformation techniques have made in Brachypodium biology as well as how Brachypodium could be used to determine the factors that might contribute to transformation efficiency. In particular, we highlight the latest research on the mechanisms that govern the gradual loss of embryogenic potential in a tissue culture and propose using B. distachyon as a model for other recalcitrant monocots.

scholarly journals Restoring the natural tropism of AAV2 vectors for human liver

2020 ◽  
Vol 12 (560) ◽  
pp. eaba3312
Author(s):  
Marti Cabanes-Creus ◽  
Claus V. Hallwirth ◽  
Adrian Westhaus ◽  
Boaz H. Ng ◽  
Sophia H.Y. Liao ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Tissue Culture ◽  
Human Liver ◽  
Xenograft Model ◽  
Human Hepatocytes ◽  
Targeted Gene Therapy ◽  
Tissue Culture Propagation ◽  
Made In

Recent clinical successes in gene therapy applications have intensified interest in using adeno-associated viruses (AAVs) as vectors for therapeutic gene delivery. Although prototypical AAV2 shows robust in vitro transduction of human hepatocyte–derived cell lines, it has not translated into an effective vector for liver-directed gene therapy in vivo. This is consistent with observations made in Fah−/−/Rag2−/−/Il2rg−/− (FRG) mice with humanized livers, showing that AAV2 functions poorly in this xenograft model. Here, we derived naturally hepatotropic AAV capsid sequences from primary human liver samples. We demonstrated that capsid mutations, likely acquired as an unintentional consequence of tissue culture propagation, attenuated the intrinsic human hepatic tropism of natural AAV2 and related human liver AAV isolates. These mutations resulted in amino acid changes that increased binding to heparan sulfate proteoglycan (HSPG), which has been regarded as the primary cellular receptor mediating AAV2 infection of human hepatocytes. Propagation of natural AAV variants in vitro showed tissue culture adaptation with resulting loss of tropism for human hepatocytes. In vivo readaptation of the prototypical AAV2 in FRG mice with a humanized liver resulted in restoration of the intrinsic hepatic tropism of AAV2 through decreased binding to HSPG. Our results challenge the notion that high affinity for HSPG is essential for AAV2 entry into human hepatocytes and suggest that natural AAV capsids of human liver origin are likely to be more effective for liver-targeted gene therapy applications than culture-adapted AAV2.

Effect Of Tissue Culture Irradiation On Orientation Of Cytoskeletal Fibers In Migrating Sheets Of Endothelial Cells

1981 ◽  
Author(s):  
A I Gotlieb
Keyword(s):  
Tissue Culture ◽  
Endothelial Cells ◽  
Immunofluorescence Microscopy ◽  
Fiber Bundles ◽  
Front Edge ◽  
Wound Edge ◽  
System A ◽  
Reduce Cell Proliferation ◽  
Made In

Irradiation of endothelial cells (EC) in tissue culture has been used to markedly reduce cell proliferation in order to study the effects of substances on the regulation of EC migration. Since irradiated EC (IRR-EC) migrate the same distance as do non-irradiated EC (NIRR-EC) over periods of up to six days, it is assumed that EC migration is not effected by irradiation. The purpose of this study was to examine the effect of irradiation on the cytoskeleton of migrating EC. The in-vitro experimental wound technique was used as a model system. A linear wound was made in confluent cultures of porcine thoracic aortic EC. EC were observed to migrate into the wound as a cohesive sheet of cells with only a few free unattached cells being present along the front edge of the wound. Cultures were irradiated one hour before wounding with 1500 rads. NIRR-EC migrated the same distance as did the IRR-EC although the latter were much flatter and each IRR-EC covered a larger surface than did the NIRR-EC. The orientation of cytoskeletal fiber bundles localized by immunofluorescence microscopy using antisera produced against electrophoretically purified porcine uterine myosin and chicken gizzard tropomyosin were different when comparing migrating IRR and NIRR-EC in the first row of the endothelial sheet. At 44 hours after wounding myosin localization showed that the main myosin stained fibers in 89% of the IRR-EC were roughly parallel to the wound edge while 11% were roughtly perpendicular to the wound edge. In NIRR-EC the figures were 54% and 46% respectively. Tropomyosin localization showed a similar diffence, 73% and 27% in IRR-EC and 45% and 55% in NIRR-EC. The data shows that there are differences in the extent of spreading of and in the orientation of cytoskeletal fiber bundles in migrating IRR-EC. These differences may reflect different cytoskeletal processes involved during migration of IRR and NIRR-EC.

scholarly journals Cell Wall Epitopes and Endoploidy as Reporters of Embryogenic Potential in Brachypodium Distachyon Callus Culture

2018 ◽  
Vol 19 (12) ◽  
pp. 3811 ◽  
Author(s):  
Alexander Betekhtin ◽  
Magdalena Rojek ◽  
Katarzyna Nowak ◽  
Artur Pinski ◽  
Anna Milewska-Hendel ◽  
...  
Keyword(s):  
Baby Boom ◽  
Brachypodium Distachyon ◽  
Plant Biotechnology ◽  
Immunohistochemical Analysis ◽  
Callus Cultures ◽  
Zygotic Embryos ◽  
Transcript Accumulation ◽  
Embryogenic Potential ◽  
Time Flow

Effective regeneration of callus tissue into embryos and then into whole plants is essential for plant biotechnology. The embryonic potential is often low and can further decrease with time in culture, which limits the utilisation of calli for transformation procedures and in vitro propagation. In this study, we show that the loss of embryogenic potential in callus cultures of Brachypodium distachyon is progressive over time. Flow cytometry analyses indicated endoploidy levels increased in 60- and 90-day-old calli with effective loss of the 2C DNA content peak in the latter. Analysis of indolic compounds content revealed a decrease in 60- and 90-day-old calli compared to either freshly isolated explants or 30-day-old calli. Immunohistochemical analysis revealed a decrease in arabinogalactan proteins (AGP) signal with the time of culture, but extensin (EXT) epitopes either increased (JIM12 epitopes) or decreased (JIM11 epitopes). The transcript accumulation levels of AGPs and EXTs confirmed these results, with most of AGP and EXT transcripts gradually decreasing. Some chimeric EXT transcripts significantly increased on the 30th day of culture, perhaps because of an increased embryogenic potential. Selected somatic embryogenesis-related genes and cyclins demonstrated a gradual decrease of transcript accumulation for YUCCA (YUC), AINTEGUMENTA-LIKE (AIL), BABY BOOM (BBM), and CLAVATA (CLV3) genes, as well as for most of the cyclins, starting from the 30th day of culture. Notably, WUSCHEL (WUS) transcript was detectable only on the 30th and 60th day and was not detectable in the zygotic embryos and in 90-day-old calli.

scholarly journals Progress in Tissue Culture and Genetic Transformation of Oil Palm: An Overview

2019 ◽  
Vol 20 (21) ◽  
pp. 5353 ◽  
Author(s):  
Rajesh Yarra ◽  
Longfei Jin ◽  
Zhihao Zhao ◽  
Hongxing Cao
Keyword(s):  
Tissue Culture ◽  
Genetic Transformation ◽  
Oil Palm ◽  
Large Scale ◽  
Elaeis Guineensis ◽  
Microprojectile Bombardment ◽  
Oil Consumption ◽  
Transformation Techniques ◽  
Significant Research

Oil palm (Elaeis guineensis, Jacq.) is a prominent vegetable-oil-yielding crop. Cultivating high-yielding oil palm with improved traits is a pre-requisite to meet the increasing demands of palm oil consumption. However, tissue culture and biotechnological approaches can resolve these concerns. Over the past three decades, significant research has been carried out to develop tissue culture and genetic transformation protocols for oil palm. Somatic embryogenesis is an efficient platform for the micropropagation of oil palm on a large scale. In addition, various genetic transformation techniques, including microprojectile bombardment, Agrobacterium tumefaciens mediated, Polyethylene glycol mediated mediated, and DNA microinjection, have been developed by optimizing various parameters for the efficient genetic transformation of oil palm. This review mainly emphasizes the methods established for in vitro propagation and genetic transformation of oil palm. Finally, we propose the application of the genome editing tool CRISPR/Cas9 to improve the various traits in this oil yielding crop.

scholarly journals How to Make Living Viral Tattoos

Leonardo ◽  
2011 ◽  
Vol 44 (2) ◽  
pp. 164-165
Author(s):  
Tagny Duff ◽  
Jill Muhling ◽  
Maria Grade Godinho ◽  
Stuart Hodgetts
Keyword(s):  
Tissue Culture ◽  
Open Source ◽  
Human Skin ◽  
Source Model ◽  
Technical Paper ◽  
Cell And Tissue Culture ◽  
Pig Skin ◽  
Materials Used ◽  
Made In

Living Viral Tattoos (2008) is a research-creation project featuring the development of sculptures made in vitro. The creation of tattoos in the form of blue ‘bruises’ on pig skin and donated human skin was made using retroviruses, cell and tissue culture and immunohistochemical stains. This technical paper presents the protocols created and materials used in the project with the intention of contributing to an open source model for the development of wetware and biological art processes.

RAPID IN VITRO PROPAGATION OF WESTERN GHATS CULTIVAR - COLOCASIA ESCULENTA FOR THE HIGH FREQUENCY OF PLANTLETS

2019 ◽  
Vol 7 (2) ◽  
Author(s):  
Jyothi R ◽  
Srinivasa Murthy K M ◽  
Hossein . ◽  
Veena .
Keyword(s):  
Tissue Culture ◽  
Wide Distribution ◽  
Colocasia Esculenta ◽  
Limiting Factor ◽  
Promising Alternative ◽  
Rapid Multiplication ◽  
Medical Plant ◽  
Total Fat ◽  
Planting Materials

Colocasia esculenta is commonly known as Taro, it is referred to as cocoyam in Nigeria. They are cherished for their rich taste, nutritional and medicinal properties. Every 100 g of taro corms possess 112 Kcal, 26.46 g carbohydrate, 1.50 g protein, 0.20 g total fat and 4.1g fiber (USDA National Nutrient Data Base). Besides its nutritional value, taro is used as a medical plant and provides bioactive compounds used as an anti-cancer drugs. Traditionally, cocoyams are vegetative propagated from tuber fragments, a practice that encourages pathogen distribution. Colocasia esculenta is a widely distributed food crop in the humid tropics and subtropics. Despite of its wide distribution, Taro plants are commonly infected with DsMV and other pathogens. This virus induces conspicuous mosaic, malformation, dwarfing or feathering on leaves in taro. As the results of infection, it reduces the quality and yield of taro production greatly. This virus is thus considered as a major limiting factor in the production of taro. Here plays the importance of  tissue culture plays a major role in producing the disease resistant plants round the year with high quality. For rapid multiplication and production of quality planting materials, tissue culture technology offers promising alternative compared to the traditional production methods. KEYWORDS: Colocasia esculenta, Virus, Pathogens, Conventional propagation, Micropropagation, Yield, Rapid multiplication, Quality

STUDIES ON THE GROWTH OF EXPERIMENTAL OVARIAN TUMOURS IN TISSUE CULTURE

1959 ◽  
Vol XXXII (I) ◽  
pp. 41-53 ◽  
Author(s):  
Stig Kullander ◽  
Bengt Källén
Keyword(s):  
Tissue Culture ◽  
Granulosa Cell ◽  
In Vitro Study ◽  
Vitro Study ◽  
Ovarian Tumours

ABSTRACT An in vitro study has been made of experimentally produced rat ovarian tumours of different age, paying particular attention to tumour reaction to crystallized steroids. Tumours of two histological structures were found: granulosa cell – luteoma tumours and arrhenoblastoma tumours. Both types grew in vitro and pictures of their cell appearance are given. The former type gave the best growth, and the endocrine studies were restricted to this type. The steroids tested (androsterone, oestrone, progesterone) all had an arresting effect in certain cases. This effect is not an unspecific, toxic one. The different tumours react to different extents, some being completely unaffected.

Potential of Nanoparticles in Combating Candida Infections

2019 ◽  
Vol 16 (5) ◽  
pp. 478-491 ◽  
Author(s):  
Faizan Abul Qais ◽  
Mohd Sajjad Ahmad Khan ◽  
Iqbal Ahmad ◽  
Abdullah Safar Althubiani
Keyword(s):  
Targeted Delivery ◽  
Recent Progress ◽  
Antifungal Agents ◽  
Fungal Infections ◽  
Fold Increase ◽  
Antifungal Drugs ◽  
Low Toxicity ◽  
Candida Infections ◽  
Made In

Aims: The aim of this review is to survey the recent progress made in developing the nanoparticles as antifungal agents especially the nano-based formulations being exploited for the management of Candida infections. Discussion: In the last few decades, there has been many-fold increase in fungal infections including candidiasis due to the increased number of immunocompromised patients worldwide. The efficacy of available antifungal drugs is limited due to its associated toxicity and drug resistance in clinical strains. The recent advancements in nanobiotechnology have opened a new hope for the development of novel formulations with enhanced therapeutic efficacy, improved drug delivery and low toxicity. Conclusion: Metal nanoparticles have shown to possess promising in vitro antifungal activities and could be effectively used for enhanced and targeted delivery of conventionally used drugs. The synergistic interaction between nanoparticles and various antifungal agents have also been reported with enhanced antifungal activity.

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