scholarly journals Usutu Virus Infection of Embryonated Chicken Eggs and a Chicken Embryo-Derived Primary Cell Line

Viruses ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 531 ◽  
Author(s):  
Emna Benzarti ◽  
José Rivas ◽  
Michaël Sarlet ◽  
Mathieu Franssen ◽  
Nassim Moula ◽  
...  
Keyword(s):  
Immune Response ◽  
Cell Line ◽  
Chicken Embryo ◽  
Vaccine Development ◽  
Chorioallantoic Membrane ◽  
Virus Production ◽  
Primary Cell ◽  
Usutu Virus ◽  
Primary Cell Line ◽  
The Future

Usutu virus (USUV) is a mosquito-borne flavivirus, closely related to the West Nile virus (WNV). Similar to WNV, USUV may cause infections in humans, with occasional, but sometimes severe, neurological complications. Further, USUV can be highly pathogenic in wild and captive birds and its circulation in Europe has given rise to substantial avian death. Adequate study models of this virus are still lacking but are critically needed to understand its pathogenesis and virulence spectrum. The chicken embryo is a low-cost, easy-to-manipulate and ethically acceptable model that closely reflects mammalian fetal development and allows immune response investigations, drug screening, and high-throughput virus production for vaccine development. While former studies suggested that this model was refractory to USUV infection, we unexpectedly found that high doses of four phylogenetically distinct USUV strains caused embryonic lethality. By employing immunohistochemistry and quantitative reverse transcriptase-polymerase chain reaction, we demonstrated that USUV was widely distributed in embryonic tissues, including the brain, retina, and feather follicles. We then successfully developed a primary cell line from the chorioallantoic membrane that was permissive to the virus without the need for viral adaptation. We believe the future use of these models would foster a significant understanding of USUV-induced neuropathogenesis and immune response and allow the future development of drugs and vaccines against USUV.

scholarly journals DIPG-50. CHROMATIN IMMUNOPRECIPITATION OF DIFFUSE INTRINSIC PONTINE GLIOMA TUMOR TISSUE IS FEASIBLE AND SHOW DIFFERENT ENRICHMENT COMPARED TO PRIMARY CELL LINE

2018 ◽  
Vol 20 (suppl_2) ◽  
pp. i59-i59
Author(s):  
Yong Yean Kim ◽  
Jaclyn Andricovich ◽  
Sridevi Yadavilli ◽  
Madhuri Kambhampati ◽  
Alexandros Tzatsos ◽  
...  
Keyword(s):  
Cell Line ◽  
Chromatin Immunoprecipitation ◽  
Tumor Tissue ◽  
Diffuse Intrinsic Pontine Glioma ◽  
Primary Cell ◽  
Pontine Glioma ◽  
Primary Cell Line ◽  
Glioma Tumor

scholarly journals Modelling of pancreatic cancer biology: transcriptomic signature for 3D PDX-derived organoids and primary cell line organoid development

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Shannon R. Nelson ◽  
Chenxi Zhang ◽  
Sandra Roche ◽  
Fiona O’Neill ◽  
Niall Swan ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cell Line ◽  
Cancer Biology ◽  
Primary Cell ◽  
Primary Cell Line ◽  
Transcriptomic Signature

scholarly journals Differential splicing of human adenovirus 5 E1A RNA expressed in cis versus in trans

2020 ◽  
Author(s):  
Drayson Graves ◽  
Nikolas Akkerman ◽  
Scott Bachus ◽  
Peter Pelka
Keyword(s):  
Gene Therapy ◽  
Cell Line ◽  
Viral Genome ◽  
Vaccine Development ◽  
Virus Production ◽  
Human Adenovirus ◽  
Virus Growth ◽  
293 Cells ◽  
In Trans ◽  
In Cis

Human adenovirus (HAdV) is used extensively as a vector for gene delivery for a variety of purposes, including gene therapy and vaccine development. Most adenoviral vectors used for these approaches have the early region 1 (E1) deleted, which is complemented by the cell line. Most commonly these are 293 cells for HAdV serotype 2 or 5. The 293 cells have the left end of HAdV5 integrated into chromosome 19 and express the E1 genes and protein IX. We observed that viruses deleted for E1 region often grow poorly on 293 cells when compared to E1 wild-type viruses. Therefore, we investigated whether this is caused by splicing differences between E1A provided by the cell line, or in trans; and that provided by the infecting viral genome, or in cis. We observed that E1A RNA that was expressed from the genome of 293 cells was spliced differently during infection with an E1A-deleted dl312 virus, versus the same cells infected with dl309 or wt300. Importantly, 293 cells were not able to fully complement the late E1A transcripts, specifically 11S, 10S, and 9S that express E1A217R, E1A171R, and E1A55R isoforms respectively. We observed that these splicing differences likely arise due to different sub-nuclear localization of E1A RNA. E1A RNA expressed from the viral genome was localized to viral replication centers, while E1A RNA expressed from the cell’s genome was not. This loss of the late E1A mRNAs and their associated proteins impacts viral growth, gene expression, and protein levels. Complementation of the late E1A mRNAs in 293 cells restored some of the observed growth defect with dl312 and resulted in higher virus growth. IMPORTANCE Human adenovirus has become an important tool for medicine and research, and 293 cells and various other similar cell lines are used extensively for virus production where high viral yields are important. Such complementing cell lines are used for production of viral vectors and vaccines, which often have deletions and replacements in various viral genes. Deletions in essential genes, such as E1, are often complemented by the cell line that is used for virus propagation in trans. Here we show that even complete genetic complementation of a viral gene does not result in full protein complementation, which compromises virus growth. This is particularly important where high viral yields are crucial, such as in virus production for vaccine development or gene therapy.

Morphology, cell viability, karyotype, expression of surface markers and plasticity of three human primary cell line cultures before and after the cryostorage in LN2 and GN2

Cryobiology ◽  
2015 ◽  
Vol 70 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Alberto Del Pino ◽  
Gertrudis Ligero ◽  
María B. López ◽  
Héctor Navarro ◽  
Jose A. Carrillo ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Line ◽  
Primary Cell ◽  
Surface Markers ◽  
Primary Cell Line ◽  
Before And After

scholarly journals PSVIII-17 In vitro and in vivo investigations of effects of natural phytogenic compounds on muscle cell gene expressions and growth performance and carcass composition of finishing pigs

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 303-304
Author(s):  
Y-J Kim ◽  
J-H Kim ◽  
J-W Kim ◽  
S-H Kim ◽  
J -S Eun ◽  
...  
Keyword(s):  
Growth Performance ◽  
Cell Line ◽  
Muscle Cell ◽  
Muscle Development ◽  
Fat Deposition ◽  
Feed Additives ◽  
Primary Cell ◽  
Synthetic Compounds ◽  
Primary Cell Line

Abstract For lean meat production in swine industry, some synthetic compounds have been applied as feed additives in finishing swine feeds. However, this practice raises public concern due to potential imbalance of innate hormones coupled with a possible residue of the synthetic compounds in meat products after ingestion. In this study, we investigated natural phytogenic compounds (NPC) from fruit peel consisted of mainly ursolic and maslinic acids to enhance muscle development but decrease fat deposition in swine. To test effects on cell line, NPC was treated in the porcine primary cell line obtained from muscle tissue of piglets at approximately 6.5 kg of body weight and NIH-3T3-L1 cell line from mouse adipose tissue cell (Experiment 1). In the porcine primary cell line, immunofluorescence measurement indicated that myogenin expression increased at 20%, while the genes responsible for muscle development increased RNA abundance in MyoD at 20%, Mrf4 at 31%, PAX3 at 19%, and PAX7 at 16% observed in myotube development from myoblast when treated with NPC (P < 0.05). In NIH-3T3L1 cell, NPC treatment increased suppression on lipid accumulation at 40% (P < 0.05). In a follow-up in vivo investigation in a completely randomized design (Experiment 2), the same NPC tested in Experiment 1 at 0.53% dry matter (DM) did not affect intake of DM, whereas it tended to increase average daily gain compared with control without NPC supplementation (1.03 vs. 0.99; P = 0.08), leading to a tendency to increase gain-to-feed ratio (0.338 vs. 0.316; P = 0.12). While NPC supplementation did not influence hot carcass weight, animals treated with NPC decreased backfat thickness compared to those fed control (23.5 vs. 24.8 mm; P < 0.01). In summary, NPC from fruit improved growth performance and carcass trait due to their impacts on muscle cell development and fat deposition.

scholarly journals Drug Repositioning Screen on a New Primary Cell Line Identifies Potent Therapeutics for Glioblastoma

2020 ◽  
Vol 14 ◽  
Author(s):  
Filiz Senbabaoglu ◽  
Ali Cenk Aksu ◽  
Ahmet Cingoz ◽  
Fidan Seker-Polat ◽  
Esra Borklu-Yucel ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Drug Repositioning ◽  
Morphological Characteristics ◽  
Genetic Alterations ◽  
Primary Cell ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Primary Cell Line

Glioblastoma is a malignant brain cancer with limited treatment options and high mortality rate. While established glioblastoma cell line models provide valuable information, they ultimately lose most primary characteristics of tumors under long-term serum culture conditions. Therefore, established cell lines do not necessarily recapitulate genetic and morphological characteristics of real tumors. In this study, in line with the growing interest in using primary cell line models derived from patient tissue, we generated a primary glioblastoma cell line, KUGBM8 and characterized its genetic alterations, long term growth ability, tumor formation capacity and its response to Temozolomide, the front-line chemotherapy utilized clinically. In addition, we performed a drug repurposing screen on the KUGBM8 cell line to identify FDA-approved agents that can be incorporated into glioblastoma treatment regimen and identified Topotecan as a lead drug among 1,200 drugs. We showed Topotecan can induce cell death in KUGBM8 and other primary cell lines and cooperate with Temozolomide in low dosage combinations. Together, our study provides a new primary cell line model that can be suitable for both in vitro and in vivo studies and suggests that Topotecan can offer promise as a therapeutic approach for glioblastoma.

scholarly journals Synthesis and biological investigation of (+)-3-hydroxymethylartemisinin

2019 ◽  
Vol 15 ◽  
pp. 567-570
Author(s):  
Toni Smeilus ◽  
Farnoush Mousavizadeh ◽  
Johannes Krieger ◽  
Xingzhao Tu ◽  
Marcel Kaiser ◽  
...  
Keyword(s):  
Cell Line ◽  
Mechanism Of Action ◽  
Antimalarial Activity ◽  
Primary Cell ◽  
Biological Investigation ◽  
Artemisinin Derivatives ◽  
L6 Cells ◽  
Skeletal Myoblasts ◽  
Primary Cell Line

Herein, we describe a biomimetic entry to (+)-3-hydroxymethylartemisinin (2) as well as to the artemisinin derivatives (+)-3-hydroxymethyl-9-desmethylartemisinin (16) and (+)-3-hydroxymethyl-9-epi-artemisinin (18), starting from the known and readily available chiral aldehyde 3 and alkyne 4. Subsequently, the synthesized compounds have been evaluated for their antimalarial activity against the drug-sensitive P. falciparum NF54 strain. All of them were inactive. In addition, they did not show any toxicity against L6 cells (a primary cell line derived from rat skeletal myoblasts). These results contribute to a better understanding of artemisinins mechanism of action.

scholarly journals Individualized Medicine for Renal Cell Carcinoma: Establishment of Primary Cell Line Culture from Surgical Specimens

2008 ◽  
Vol 22 (10) ◽  
pp. 2361-2366 ◽  
Author(s):  
Fernando J. Kim ◽  
Adriano Campagna ◽  
Lakshmipathi Khandrika ◽  
Sweaty Koul ◽  
Seok-Soo Byun ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Cell Line ◽  
Renal Cell ◽  
Individualized Medicine ◽  
Primary Cell ◽  
Primary Cell Line
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