The Cellular Substrate: A Very Important Requirement for Baculovirus in vitro Replication

1984 ◽  
Vol 39 (9-10) ◽  
pp. 993-1002 ◽  
Author(s):  
Herbert G. Miltenburger ◽  
Werner L. Naser ◽  
Jeanne P. Harvey ◽  
Jürg Huber ◽  
Alois M. Huger
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cydia Pomonella ◽  
Codling Moth ◽  
Primary Cell ◽  
Insect Species ◽  
In Vitro Replication ◽  
Primary Cell Lines ◽  
First Time

Abstract We established more than 200 primary cell lines of Cydia pomonella (codling moth). 81 of them were selected and screened for replication of two baculoviruses (from two different subgroups): the Choristoneura murinana NPV and the Cydia pomonella GV. Although all these cell lines had been derived from the same insect species, they varied largely in their response to challenge with the NPV. Most of them showed CPE or produced different amounts of poly-hedra. Interestingly, we also found a few cell lines that were permissive for GV replication. To our knowledge this is the first time that GV replication in cell lines has been obtained. Our results show that cell line properties are most important for baculovirus in vitro replication.

scholarly journals In vitro and in vivo effect of human lactoferrin on glioblastoma growth

2015 ◽  
Vol 123 (4) ◽  
pp. 1026-1035 ◽  
Author(s):  
Antonietta Arcella ◽  
Maria Antonietta Oliva ◽  
Sabrina Staffieri ◽  
Silvia Aalberti ◽  
Giovanni Grillea ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Nude Mice ◽  
Tumor Size ◽  
Primary Cell ◽  
Continuous Cell Line ◽  
Human Lactoferrin ◽  
Primary Cell Lines

OBJECT Human lactoferrin (HLF) is a natural protein with antitumor activity. The aim of this study was to investigate the effects of HLF alone and in combination with temozolomide (TMZ), a conventional chemotherapeutic, on human glioblastoma (GBM) cells. METHODS The authors cultured fresh human primary cell lines NMD and FN and the continuous cell line U87MG to evaluate proliferation in the presence of HLF alone at different doses (1, 10, and 100 mg/ml, and 1 mg/ml) and in combination with TMZ. In in vivo experiments they assessed tumor size reduction in CD1 nude mice carrying an orthotopic GBM xenograft and orally treated with HLF. RESULTS Lactoferrin causes growth inhibition in the NMD and FN primary cell lines and in the U87MG continuous cell line. This inhibition seemed to be modulated by the downregulation of cyclin D1 and D4. Western blot and fluorescence-activated cell sorting analysis showed inhibition of the cell cycle in G0/G1 and G2 phases. When administered in nude mice, HLF (60 mg/kg/day) decreased tumor size about 30%, as shown in both histological analyses and high-field brain MRI. Administration of HLF with TMZ enhanced the effect of chemotherapy both in vitro and in vivo. CONCLUSIONS This study demonstrated that HLF can inhibit GBM cell growth, suggesting that this nontoxic substance may have a role in potentiating the effect of current TMZ treatment of GBM.

scholarly journals Analysis of a transgenic mouse containing simian virus 40 and v-myc sequences.

1985 ◽  
Vol 5 (4) ◽  
pp. 642-648 ◽  
Author(s):  
J A Small ◽  
D G Blair ◽  
S D Showalter ◽  
G A Scangos
Keyword(s):  
Cell Lines ◽  
Choroid Plexus Papilloma ◽  
Simian Virus 40 ◽  
Soft Agar ◽  
Simian Virus ◽  
T Antigen ◽  
Primary Cell ◽  
Tissue Samples ◽  
Primary Cell Lines

Two plasmids, one containing the simian virus 40 (SV40) genome and the mouse metallothionein I gene and one containing the v-myc gene of avian myelocytomatosis virus MC29, were coinjected into mouse embryos. Of the 13 surviving mice, one, designated M13, contained both myc and SV40 sequences. This mouse developed a cranial bulge identified as a choroid plexus papilloma at 13 weeks and was subsequently sacrificed; tissue samples were taken for further analysis. Primary cell lines derived from these tissues contained both myc and SV40 DNA. No v-myc mRNA could be detected, although SV40 mRNA was present in all of the cell lines tested. T antigen also was expressed in all of the cell lines analyzed. These data suggest that SV40 expression was involved in the abnormalities of mouse M13 and was responsible for the transformed phenotype of the primary cell lines. Primary cell lines from this mouse were atypical in that the population rapidly became progressively more transformed with time in culture based on the following criteria: morphology, growth rate, and the ability to grow in soft agar and in serum-free medium. The data also suggest that factors present in the mouse regulated the ability of SV40 to oncogenically transform most cells and that in vitro culture of cells allowed them to escape those factors.

In vitro effects of erythromycin and florfenicol on primary cell lines of Unio crassus and Cyprinus carpio

2021 ◽  
Author(s):  
Pınar Arslan ◽  
Begum Yurdakok-Dikmen ◽  
Saniye Cevher Ozeren ◽  
Ozgur Kuzukiran ◽  
Ayhan Filazi
Keyword(s):  
Cell Lines ◽  
Cyprinus Carpio ◽  
Primary Cell ◽  
Primary Cell Lines ◽  
In Vitro Effects

scholarly journals Aminolevulinic Acid-Mediated Photodynamic Therapy of Human Meningioma: An in Vitro Study on Primary Cell Lines

2015 ◽  
Vol 16 (12) ◽  
pp. 9936-9948 ◽  
Author(s):  
Mustafa El-Khatib ◽  
Carolin Tepe ◽  
Brigitte Senger ◽  
Maxine Dibué-Adjei ◽  
Markus Riemenschneider ◽  
...  
Keyword(s):  
Photodynamic Therapy ◽  
Cell Lines ◽  
Aminolevulinic Acid ◽  
In Vitro Study ◽  
Vitro Study ◽  
Primary Cell ◽  
Human Meningioma ◽  
Primary Cell Lines

scholarly journals Molluscan cells in culture: primary cell cultures and cell lines

2013 ◽  
Vol 91 (6) ◽  
pp. 391-404 ◽  
Author(s):  
T.P. Yoshino ◽  
U. Bickham ◽  
C.J. Bayne
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cell Cultures ◽  
Snail Host ◽  
Primary Cell ◽  
Neoplastic Disease ◽  
Freshwater Bivalve ◽  
Primary Cell Cultures ◽  
Organ Systems

In vitro cell culture systems from molluscs have significantly contributed to our basic understanding of complex physiological processes occurring within or between tissue-specific cells, yielding information unattainable using intact animal models. In vitro cultures of neuronal cells from gastropods show how simplified cell models can inform our understanding of complex networks in intact organisms. Primary cell cultures from marine and freshwater bivalve and gastropod species are used as biomonitors for environmental contaminants, as models for gene transfer technologies, and for studies of innate immunity and neoplastic disease. Despite efforts to isolate proliferative cell lines from molluscs, the snail Biomphalaria glabrata (Say, 1818) embryonic (Bge) cell line is the only existing cell line originating from any molluscan species. Taking an organ systems approach, this review summarizes efforts to establish molluscan cell cultures and describes the varied applications of primary cell cultures in research. Because of the unique status of the Bge cell line, an account is presented of the establishment of this cell line, and of how these cells have contributed to our understanding of snail host – parasite interactions. Finally, we detail the difficulties commonly encountered in efforts to establish cell lines from molluscs and discuss how these difficulties might be overcome.

scholarly journals AAV9 delivering a modified human Mullerian inhibiting substance as a gene therapy in patient-derived xenografts of ovarian cancer

2015 ◽  
Vol 112 (32) ◽  
pp. E4418-E4427 ◽  
Author(s):  
David Pépin ◽  
Amanda Sosulski ◽  
Lihua Zhang ◽  
Dan Wang ◽  
Vinod Vathipadiekal ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Gene Therapy ◽  
Cell Lines ◽  
Receptor Expression ◽  
Primary Cell ◽  
Blood Concentrations ◽  
Müllerian Inhibiting Substance ◽  
Primary Cell Lines ◽  
Mullerian Inhibiting Substance

To improve ovarian cancer patient survival, effective treatments addressing chemoresistant recurrences are particularly needed. Mullerian inhibiting substance (MIS) has been shown to inhibit the growth of a stem-like population of ovarian cancer cells. We have recently engineered peptide modifications to human MIS [albumin leader Q425R MIS (LRMIS)] that increase production and potency in vitro and in vivo. To test this novel therapeutic peptide, serous malignant ascites from highly resistant recurrent ovarian cancer patients were isolated and amplified to create low-passage primary cell lines. Purified recombinant LRMIS protein successfully inhibited the growth of cancer spheroids in vitro in a panel of primary cell lines in four of six patients tested. Adeno-associated virus (AAV) -delivered gene therapy has undergone a clinical resurgence with a good safety profile and sustained gene expression. Therefore, AAV9 was used as a single i.p. injection to deliver LRMIS to test its efficacy in inhibiting growth of palpable tumors in patient-derived ovarian cancer xenografts from ascites (PDXa). AAV9-LRMIS monotherapy resulted in elevated and sustained blood concentrations of MIS, which significantly inhibited the growth of three of five lethal chemoresistant serous adenocarcinoma PDXa models without signs of measurable or overt toxicity. Finally, we tested the frequency of MIS type II receptor expression in a tissue microarray of serous ovarian tumors by immunohistochemistry and found that 88% of patients bear tumors that express the receptor. Taken together, these preclinical data suggest that AAV9-LRMIS provides a potentially well-tolerated and effective treatment strategy poised for testing in patients with chemoresistant serous ovarian cancer.

scholarly journals TMOD-05. GENOME-WIDE DNA METHYLATION PROFILE: A POWERFUL STRATEGY TO RECAPITULATE HETEROGENEITY OF PEDIATRIC BRAIN TUMORS IN PRIMARY CELL LINES

2021 ◽  
Vol 23 (Supplement_1) ◽  
pp. i36-i36
Author(s):  
Lucia Pedace ◽  
Simone Pizzi ◽  
Maria Vinci ◽  
Giulia Pericoli ◽  
Giuseppina Catanzaro ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Cell Lines ◽  
Pediatric Brain Tumors ◽  
Genetic Alterations ◽  
In Vitro Models ◽  
Primary Cell ◽  
Genome Wide ◽  
Primary Cell Lines ◽  
Pediatric Brain

Abstract Background Development of in vitro models of pediatric brain tumors (pBT) is instrumental for both understanding the contributing oncogenic molecular mechanisms and identifying and testing new therapeutic strategies. Primary cell lines should be established and managed to prevent epigenetic and genetic alterations and thus recapitulating the original tumor. DNA methylation (DM) is a stable epigenetic modification, altered in cancer and recently used to classify tumors. We aim to apply DM and Copy Number Variation (CNV) profiling to characterize pBT primary cell lines and tumors. Methods We investigated 34 pBT tissues from different histology paired to 52 their derived primary cultures in both 2D and 3D conditions, as stem-cells or in serum-supplemented medium, and both short and long-terms in culture. We studied 18 additional pBT-derived cell-lines, 9 organoids, 5 commercial cell-lines, and 122 pBT tissues from the same histological categories, as controls, for a total of 240 genome-wide DM profiles. We analyzed DM and CNV profiles by using Illumina EPIC-arrays. By means of a bump hunting strategy, we identified differentially methylated regions in faithful vs unfaithful cell lines, and performed a functional characterization using over-representation analysis. Results The 69% (25/36) of cells at early passages retained genetic alteration and the same DM patterns of the original tumors, with no differences related to 2D/3D methods or the presence of serum in media. The 70% (24/34) of primary cell lines analyzed at later passages (>5 or >14 days in culture) diverged from the primary tumor, the totality of those cultured with serum. All divergent cells clustered together acquiring common deregulated epigenetic signature induced by serum culture media, 2D methods and longer time in culture. Conclusions We have shown that global DM profiles, along with CNV analysis are useful tools to detect the recapitulation of pBT-derived primary cell-lines from the original tumor. Whatever subgroups tested, our results suggest that in vitro models should be passaged as little as possible to retain the epigenetic and genetic alterations of the tumors and thus to be considered relevant for basic and translational biology.

In vitro Cytotoxic Activity of Salsola oppositifolia Desf. (Amaranthaceae) in a Panel of Tumour Cell Lines

2008 ◽  
Vol 63 (5-6) ◽  
pp. 347-354 ◽  
Author(s):  
Rosa Tundis ◽  
Monica R. Loizzo ◽  
Marco Bonesi ◽  
Federica Menichini ◽  
Giancarlo A. Statti ◽  
...  
Keyword(s):  
Cytotoxic Activity ◽  
Cell Line ◽  
Cell Lines ◽  
Amelanotic Melanoma ◽  
Strong Activity ◽  
Etoac Fraction ◽  
Tumour Cell Lines ◽  
First Time ◽  
Mcf 7

The aim of the present study was to evaluate for the first time the in vitro cytotoxic activity of fractions and isolated flavonols from Salsola oppositifolia Desf. (Amaranthaceae). The nhexane fraction demonstrated an effective cytotoxic activity on the large lung carcinoma and amelanotic melanoma cell lines with IC50 values of 19.1 μg/ml and 24.4 μg/ml, respectively. Also the dichloromethane fraction exhibited cytotoxic activity against COR-L23 (IC50 30.4 μg/ml) and C32 (IC50 33.2 μg/ml) cells, while the EtOAc fraction demonstrated a selective cytotoxic activity against MCF-7 cells (IC50 67.9 μg/ml). The major active constituents of this fraction were isorhamnetin-3-O-glucoside (1) and isorhamnetin-3-O-rutinoside (2), which showed an interesting activity against the cell line MCF-7 with IC50 values of 18.2 and 25.2 μg/ml, respectively. Compound 2 exhibited a strong activity against the hormonedependent prostate carcinoma LNCaP cell line with an IC50 of 20.5 μg/ml. Constituents of S. oppositifolia were identified by GC-MS and NMR analyses.

scholarly journals Apatinib Mesylate Inhibits the Proliferation and Metastasis of Epithelioid Malignant Peritoneal Mesothelioma In Vitro and In Vivo

2020 ◽  
Vol 10 ◽  
Author(s):  
Zhi-Ran Yang ◽  
Zhi-Gao Chen ◽  
Xue-Mei Du ◽  
Yan Li
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Peritoneal Mesothelioma ◽  
Primary Cell ◽  
Malignant Peritoneal Mesothelioma ◽  
Primary Cell Lines ◽  
Proliferation And Metastasis ◽  
Pdx Models

ObjectiveMalignant peritoneal mesothelioma (MPM) is a rare malignancy with few effective molecular therapies. In this study, we evaluated the anti-tumor activity and safety of apatinib, a vascular endothelial growth factor receptor 2 inhibitor, in MPM in vitro and in vivo.MethodsWe established several patient-derived xenograft (PDX) models and primary cell lines of MPM. The cell lines were used to study the effects of apatinib on proliferation, cell cycle, migration, and apoptosis by CCK8, flow cytometry, wound-healing, Transwell, DAPI staining, and caspase-3 assays, respectively. For in vivo study, apatinib was delivered by gastric gavage into PDX models, and then efficacy and toxicity were determined by experimental peritoneal cancer index (ePCI) score and pathological examinations.ResultsOur results showed that apatinib significantly inhibited the proliferation and migration of MPM cells in vitro and induced cell cycle arrest. Studies on PDX models concurred that apatinib effectively suppressed subphrenic and liver invasions of nude mice. Moreover, histopathological analysis found that lymphocyte infiltration, coagulation necrosis and eosinophilic cell fragments were detected in tumor tissues after apatinib treatment. Apatinib showed no obvious effects on body mass of models and did not affect function of important organs, except for occasional focal lymphoid infiltration of liver (16.7%) and cardiac muscle (16.7%).ConclusionsWe successfully established MPM PDX models and primary cell lines, and confirmed that apatinib effectively inhibited proliferation and metastasis of MPM in vitro and in vivo study.

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