Persistent infection of Malacosoma disstria (Lepidoptera: Lasiocampidae) cell cultures with Nosema (Glugea) disstriae (Microsporida: Nosematidae)

1976 ◽  
Vol 54 (3) ◽  
pp. 336-342 ◽  
Author(s):  
S. S. Sohi ◽  
G. G. Wilson
Keyword(s):  
Cell Lines ◽  
Cell Cultures ◽  
Persistent Infection ◽  
Malacosoma Disstria

Cell lines were developed from hemocytes and ovarian tissues of Malacosoma disstria larvae that were naturally infected with Nosema disstriae. The infection was carried over into the cell cultures. N. disstriae disappeared from the ovarian cultures after several passages in vitro involving a period of more than 2 years, but hemocyte cultures are still infected after 6 years (199 passages). The spores produced in cell cultures were infectious for M. disstria larvae.

Hepatoma Cell Cultures as In Vitro Models for the Hepatotoxicity of Xenobiotics

1988 ◽  
Vol 16 (1) ◽  
pp. 32-37
Author(s):  
Margherita Ferro ◽  
Anna Maria Bassi ◽  
Giorgio Nanni
Keyword(s):  
Cell Lines ◽  
Cell Cultures ◽  
Membrane Lipids ◽  
Hepatoma Cell ◽  
Positive Control ◽  
In Vitro Models ◽  
Low Concentrations ◽  
Formation Efficiency ◽  
Dose Dependent

Two hepatoma cell cultures were examined as in vitro models to be used in genotoxicity and cytotoxicity tests without the addition of bioactivating enzymes. The MH1C1, and HTC hepatoma lines were used in this study to establish their sensitivity to a number of xenobiotics, namely, cyclophosphamide (CP), the classical positive control in bioactivation tests; benzaldehyde (BA), a short-chain aldehyde; and 4-hydroxynonenal (HNE), a major toxic end-product of the peroxidative degradation of cell membrane lipids. As a first approach, we compared the following cytotoxicity tests: release of lactate dehydrogenase (LDH), and colony formation efficiency (CF). Colony-forming cells were exposed to the drugs according to different procedures, before or after the anchorage phase. The leakage of LDH into the medium following exposure of both cell lines to HNE, CP and BA for up to 24 hours was found not to be a good index of cytotoxicity. A better indicator of cytotoxicity was CF, as evaluated by exposure of the cells 24 hours after seeding. The effects were detectable at very low concentrations, corresponding to 10, 90 and 100μM for HNE, CP and BA, respectively. The impairment of CF efficiency was dose-dependent and time-dependent, and several differences between the two cell lines were observed.

scholarly journals Isolation and initial propagation of guinea pig adenovirus (GPAdV) in Cavia porcellus cell lines

F1000Research ◽  
2020 ◽  
Vol 8 ◽  
pp. 1597
Author(s):  
Adriana E. Kajon ◽  
Xiaoxin Li ◽  
Gabriel Gonzalez ◽  
Susan Core ◽  
Helga Hofmann-Sieber ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Cell Lines ◽  
Cell Cultures ◽  
Sequence Data ◽  
Respiratory Illness ◽  
Epithelial Cell Line ◽  
Cavia Porcellus ◽  
Diagnostic Assays ◽  
Tracheal Epithelial

Background:  The lack of adequate in vitro systems to isolate and propagate guinea pig adenovirus (GPAdV), a prevalent cause of respiratory illness of varaible severity in laboratory guinea pig colonies worldwide, has precluded its formal characterization to allow for the development of comprehensive diagnostic assays, and for the execution of complex pathogenesis and basic virology studies. Methods: Two strains of GPAdV were isolated in guinea pig (Cavia porcellus) cell cultures from frozen archival infected animal tissue originated from colony outbreaks of pneumonia in Australia and the Czech Republic in 1996. Results: Commercially available guinea pig cell lines from colorectal carcinoma (GPC-16), fetal fibroblast (104-C1) and lung fibroblast (JH4 C1), and the tracheal epithelial cell line GPTEC-T developed in this study were able to support viral infection and early propagation. Sufficient viral DNA was recovered from cell cultures to PCR-amplify and obtain sequence data for the complete hexon gene and partial DNA polymerase and penton base genes. Phylogenetic analysis for the three regions of the genome provided strong evidence confirming GPAdV as a unique species in the genus Mastadenovirus. Conclusions: This study demonstrated the feasibility of propagating GPAdV in cultures of immortalized lines of GP cells of a variety of types, thus establishing a critical foundation for the development of a robust culture platform for virus stock production and titration. The generation and analysis of whole GPAdV genome sequences will provide additional data for a comprehensive description of the genetic organization of the viral genome and for a better assessment of genetic diversity between the two isolated strains.

Variances in the Expression Profile of the EMT-Related Genes in Endometrial Cancer Lines In Vitro Study

2021 ◽  
Vol 22 ◽  
Author(s):  
Robert Nowakowski ◽  
Beniamin Grabarek ◽  
Anna Burnat-Olech ◽  
Dariusz Boroń ◽  
Monika Paul-Samojedny
Keyword(s):  
Endometrial Cancer ◽  
Expression Pattern ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cell Cultures ◽  
Expression Profile ◽  
Histological Grade ◽  
Cancer Cell Lines ◽  
Endometrial Cancer Cell

Background: This study aimed to evaluate the variances in the expression pattern of mRNAs and miRNAs related to the EMT in the Ishikawa (histological grade 1; G1), EC-1A (histological grade 2; G2), and KLE (histological grade 3; G3) cell cultures under cisplatin treatment. Methods: Endometrial cancer cell lines were exposed to 75.22 mg (an average concentration of the drug used in patients with endometrial cancer) for 12.24 and 48 hours compared to the untreated cells (control). The molecular analysis included extraction of total RNA, microarray analysis (mRNA and miRNA), RTqPCR, and the ELISA assay. Results: Out of 226 mRNAs associated with the EMT, the number of mRNAs differentially expressed in endometrial cancer cell cultures treated with cisplatin compared to a control culture was as follows: Ishikawa line - 87 mRNAs; EC-1A - 84 mRNAs; KLE - 71 mRNAs (p<0.05). The greatest changes in the Ishikawa line treated with the drug compared to the control were noticed for mRNA STAT1 TGFβ1, SMAD3, FOXO8, whereas in EC-1A, they were mRNA TGFβ1, BAMBI, SMAD4, and in KLE mRNA COL1A1, FOXO8, TGFβ1. The analysis also showed that miR-106a, miR-30d, miR-300 are common for all cell lines used in this experiment. Conclusion: Cisplatin changes the expression profile of genes associated with EMT in endometrial cancer cell lines. It seems that the expression pattern of TGFβ1 might be a promising, supplementary molecular marker of the effectiveness of cisplatin therapy. The analysis showed that miR-30d, miR-300, and miR-106a are involved in the regulation of the expression of EMT-related genes.

Persistent Infection of Human Oligodendrocytic and Neuroglial Cell Lines by Human Coronavirus 229E

1999 ◽  
Vol 73 (4) ◽  
pp. 3326-3337 ◽  
Author(s):  
Nathalie Arbour ◽  
Sophie Ekandé ◽  
Geneviève Côté ◽  
Claude Lachance ◽  
Fanny Chagnon ◽  
...  
Keyword(s):  
Nervous System ◽  
Cell Lines ◽  
Persistent Infection ◽  
Viral Antigen ◽  
Infectious Virus ◽  
Acute Infection ◽  
Viral Rna ◽  
Persistent Infections ◽  
Virus Progeny

ABSTRACT Human coronaviruses (HuCV) cause common colds. Previous reports suggest that these infectious agents may be neurotropic in humans, as they are for some mammals. With the long-term aim of providing experimental evidence for the neurotropism of HuCV and the establishment of persistent infections in the nervous system, we have evaluated the susceptibility of various human neural cell lines to acute and persistent infection by HuCV-229E. Viral antigen, infectious virus progeny and viral RNA were monitored during both acute and persistent infections. The astrocytoma cell lines U-87 MG, U-373 MG, and GL-15, as well as neuroblastoma SK-N-SH, neuroglioma H4, and oligodendrocytic MO3.13 cell lines, were all susceptible to an acute infection by HuCV-229E. The CHME-5 immortalized fetal microglial cell line was not susceptible to infection by this virus. The MO3.13 and H4 cell lines also sustained a persistent viral infection, as monitored by detection of viral antigen and infectious virus progeny. Sequencing of the S1 gene from viral RNA after ∼130 days of infection showed two point mutations, suggesting amino acid changes during persistent infection of MO3.13 cells but none for H4 cells. Thus, persistent in vitro infection did not generate important changes in the S1 portion of the viral spike protein, which was shown for murine coronaviruses to bear hypervariable domains and to interact with cellular receptor. These results are consistent with the potential persistence of HuCV-229E in cells of the human nervous system, such as oligodendrocytes and possibly neurons, and the virus’s apparent genomic stability.

scholarly journals Cytotoxic Effect of Trabectedin In Human Adrenocortical Carcinoma Cell Lines and Primary Cells

Cancers ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 928 ◽  
Author(s):  
Andrea Abate ◽  
Elisa Rossini ◽  
Sara Anna Bonini ◽  
Martina Fragni ◽  
Deborah Cosentini ◽  
...  
Keyword(s):  
Cytotoxic Activity ◽  
Cell Lines ◽  
Cell Cultures ◽  
Adrenocortical Carcinoma ◽  
Cytotoxic Effect ◽  
Alkylating Agents ◽  
Primary Cell ◽  
Primary Cell Cultures ◽  
High Cytotoxicity

Mitotane is the only drug approved for the treatment of adrenocortical carcinoma (ACC). The regimen to be added to mitotane is a chemotherapy including etoposide, doxorubicin, and cisplatin. This pharmacological approach, however, has a limited efficacy and significant toxicity. Evidence indicates that ACC seems to be sensitive to alkylating agents. Trabectedin is an anti-tumor drug that acts as an alkylating agent with a complex mechanism of action. Here, we investigated whether trabectedin could exert a cytotoxic activity in in vitro cell models of ACC. Cell viability was evaluated by MTT assay on ACC cell lines and primary cell cultures. The gene expression was evaluated by q-RT-PCR, while protein expression and localization were studied by Western blot and immunocytochemistry. Combination experiments were performed to evaluate their interaction on ACC cell line viability. Trabectedin demonstrated high cytotoxicity at sub-nanomolar concentrations in ACC cell lines and patient-derived primary cell cultures. The drug was able to reduce /β catenin nuclear localization, although it is unclear whether this effect is involved in the observed cytotoxicity. Trabectedin/mitotane combination exerted a synergic cytotoxic effect in NCI-H295R cells. Trabectedin has antineoplastic activity in ACC cells. The synergistic cytotoxic activity of trabectedin with mitotane provides the rationale for testing this combination in a clinical study.

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 Studies of Melody Efficacy Vaccine Against SARS-CoV-2 are Closing COVID-19 Pandemic Doors? Single Dose is Recommended as Therapeutic Drug and Two Doses Application as Preventive Vaccine

2021 ◽  
Keyword(s):  
Single Dose ◽  
Cell Lines ◽  
Cell Cultures ◽  
Statistical Difference ◽  
In Vitro Studies ◽  
Therapeutic Drug ◽  
In Vitro Test ◽  
Preventive Vaccine ◽  
Vitro Test

In vitro studies provide data that is important for proof-of-concept determination, function validation, and peer review manuscript preparation, FDA applications and clinical trials. Our in vitro studies of Melody efficacy as a vaccine against SARS-CoV-2 were made in five cell cultures (BEAS-2B, HUVEC, HMEC-1, HEK293 and WM266), where the dead cell number monitored by using flow cytometry. Melody’s efficacy was tested by incubating melody RpA (RNA-peptide A) and melody RpAB (RNA-peptide AB). Both incubations were pending 16 hours with melody Lethal Concentration 50 % (LC50 µg/µL), which was determinated in a toxicological assay in the five cell cultures as previously described. Our conclusion was focused to understand the statistical difference in LC50 in order to estimate the melody preventive efficacity as vaccine in “In vitro” test. In the present study, the preventive efficacy values obtained in the five cell lines differ statistically between the value of two doses when compared with a single or three doses, p=5.321. These results suggested that the application of two doses is more effective. We used the Student’s t-test with n=6 to calculate p-value according to mRNA avian coronavirus action after one, two and three doses incubations of mRNA-peptide-A (melody therapeutic) of 16 hours. Our conclusion was focused to understand the statistical difference in LC50 in order to estimate the melody therapeutic efficacity as vaccine in “In vitro” test in cell lines (BEAS-2B, HUVEC, HMEC-1, HEK293 and WM-266). Results: Melody Therapeutic Efficacity was found to be 93.82% with using a single dose. Meanwhile, Melody Preventive Efficacity was found as 92.53% with using two doses. The therapeutic efficacy values obtained in the five cell lines in this study did not vary statistically significantly between the three doses used, p**=0.004. As a result, the Melody application of a single dose is recommended as therapeutic drug and two doses application as preventive vaccine.

Obtaining and Characteristics of Plant Cell Cultures of Аjuga turkestanica (Rgl.) - Producing of Phytoexdystroides

2019 ◽  
pp. 50-56
Author(s):  
G.I. Sobolkova ◽  
T.D. Kharitonov ◽  
A.M. Nigmatullaev ◽  
Sh.Sh. Sagdullaev ◽  
D.V. Kochkin ◽  
...  
Keyword(s):  
Cell Lines ◽  
Plant Cell ◽  
Cell Cultures ◽  
Plant Cell Cultures ◽  
Suspension Cell ◽  
Wild Plant ◽  
Russian Science ◽  
Growth Cycles ◽  
Ajuga Turkestanica

Callus cultures of Ajuga turkestanica (Rgl.) Briq. (Lamiaceae) were obtained from a wild plant, and the processes of morphogenesis in these cultures were studied. Depending on the hormonal composition of media, it is possible to obtain gemmogenesis or rhizogenesis in the first 6 to 8 growth cycles; after 10-15 growth cycles, the ability of morphogenesis was lost. Suspension cell cultures were initiated from some well-growing calluses. As a result, about one hundred calluses and suspension cell lines were obtained from the Ajuga turkestanica plant cells, a number of which were characterized by intensive growth and were analyzed by the HPLC-MS method for the presence of phytoecdysteroids. In most of the investigated lines, 20-hydroxyecdysone and turkesterone were found, the content of the first being 30-50 times higher than the second (in the most productive lines, 2.0-2.5 mg/g and 0.04-0.05 mg/g dry weight, respectively). An increase in the content of phytoecdysteroids in the in vitro cultivated cells by the end of the growing cycle was observed. However, phytoecdysteroids were not found in many of the obtained cell lines. Further research is needed to clarify the reasons for the presence or absence of phytoecdysteroids in Ajuga turkestanica plant cell cultures. Ajuga turkestanica, plant cell culture, growth regulators, differentiation, gemmogenesis, rhizogenesis, phytoecdysteroids, 20-hydroxy ecdysone, turkesterone The work was financially supported by the Russian Science Foundation (№ 16-14-00126).

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