Establishment and Characterization of Eye Muscle Cell Line from Snow Trout, Schizothorax richardsonii (Gray, 1832), a Vulnerable Coldwater Fish, for In Vitro Studies

Schizothorax richardsonii, commonly called snow trout is a vulnerable coldwater fish distributed in India and adjoining countries, is an important resource as food, ornamental and game fishery. A cell line, designated as SREM-1, has been established from the eye muscle of S. richardsonii using explant method and cultured in Leibovitz’s L-15 medium. The growth studies at 20-32°C temperatures and 5-20% of fetal bovine serum (FBS) concentrations revealed that SREM-1 cells exhibited optimal growth at 28°C in L-15 medium containing 20% FBS. Cell type was confirmed primarily as epithelial in nature by immuno-phenotyping assay and was sub-cultured for >55 times since its development. The origin of the cell line was confirmed by sequencing of cytochrome oxidase c subunit I and 16S rRNA genes. SREM-1 cell line is free from mycoplasma contamination and has been cryopreserved at different passage levels with revival efficiency of 70-80% after 6 months. The SREM-1 cell line was successfully transfected and also employed for evaluating in vitro cytotoxicity against a heavy metal, mercuric chloride. The newly established cell line of this vulnerable species would be useful as a model for aquatic toxicological and transfection studies.

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Characterization of a new cell line from ornamental fish Amphiprion ocellaris (Cuvier, 1830) and its susceptibility to nervous necrosis virus

Scientific Reports ◽

10.1038/s41598-020-76807-7 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

B. S. Yashwanth ◽

Mukunda Goswami ◽

Rajendran Kooloth Valappil ◽

Dimpal Thakuria ◽

Aparna Chaudhari

Keyword(s):

Cell Line ◽

Plasmid Vector ◽

Chromosome Analysis ◽

Ornamental Fish ◽

16S Rrna Genes ◽

Rrna Genes ◽

Nervous Necrosis Virus ◽

Necrosis Virus ◽

A Cell

AbstractAmphiprion ocellaris (ocellaris clownfish) is one of the most commercially important marine ornamental fish. A cell line designated as OCF was developed for the first time from the caudal fin of this fish species. The cell line was maintained in Leibovitz’s—15 medium supplemented with 15% FBS (Fetal Bovine Serum) and was successfully subcultured up to 34 passages. The cell line was authenticated by sequencing mitochondrial cytochrome C oxidase subunit I (COI) and 16S rRNA genes. The growth rate of the OCF cell line was maximum in medium containing 20% FBS and 1% of 0.2 M NaCl at 28 °C. Chromosome analysis revealed 48 diploid chromosomes. The OCF cell line was transfected with the pMaxGFP plasmid vector with 7% efficiency and GFP expression was observed. The OCF cell line was used for testing nervous necrosis virus (NNV) susceptibility. Cytopathic effect (CPE) was observed in terms of plaque formation after virus inoculation. Nested PCR confirmed the susceptibility of the OCF cell line to NNV. The cell line was successfully cryopreserved by a slow freezing procedure at − 80 °C with a revival efficiency of 70–75%. The study revealed that the OCF cell line would be useful for virological studies. In addition, the cell line would play an important role as an in vitro tool for carrying out toxicological and biotechnological studies.

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5-Nitro-Thiophene-Thiosemicarbazone Derivatives Present Antitumor Activity Mediated by Apoptosis and DNA Intercalation

Current Topics in Medicinal Chemistry ◽

10.2174/1568026619666190621120304 ◽

2019 ◽

Vol 19 (13) ◽

pp. 1075-1091 ◽

Cited By ~ 4

Author(s):

Karla Mirella Roque Marques ◽

Maria Rodrigues do Desterro ◽

Sandrine Maria de Arruda ◽

Luiz Nascimento de Araújo Neto ◽

Maria do Carmo Alves de Lima ◽

...

Keyword(s):

Cell Cycle ◽

Antitumor Activity ◽

Cell Line ◽

Mitochondrial Membrane ◽

In Vitro Cytotoxicity ◽

Dna Intercalation ◽

Phase Cell ◽

Fluorescence Studies ◽

In Silico Studies

Background: Considering the need for the development of new antitumor drugs, associated with the great antitumor potential of thiophene and thiosemicarbazonic derivatives, in this work we promote molecular hybridization approach to synthesize new compounds with increased anticancer activity. Objective: Investigate the antitumor activity and their likely mechanisms of action of a series of N-substituted 2-(5-nitro-thiophene)-thiosemicarbazone derivatives. Methods: Methods were performed in vitro (cytotoxicity, cell cycle progression, morphological analysis, mitochondrial membrane potential evaluation and topoisomerase assay), spectroscopic (DNA interaction studies), and in silico studies (docking and molecular modelling). Results: Most of the compounds presented significant inhibitory activity; the NCIH-292 cell line was the most resistant, and the HL-60 cell line was the most sensitive. The most promising compound was LNN-05 with IC50 values ranging from 0.5 to 1.9 µg.mL-1. The in vitro studies revealed that LNN-05 was able to depolarize (dose-dependently) the mitochondrial membrane, induceG1 phase cell cycle arrest noticeably, promote morphological cell changes associated with apoptosis in chronic human myelocytic leukaemia (K-562) cells, and presented no topoisomerase II inhibition. Spectroscopic UV-vis and molecular fluorescence studies showed that LNN compounds interact with ctDNA forming supramolecular complexes. Intercalation between nitrogenous bases was revealed through KI quenching and competitive ethidium bromide assays. Docking and Molecular Dynamics suggested that 5-nitro-thiophene-thiosemicarbazone compounds interact against the larger DNA groove, and corroborating the spectroscopic results, may assume an intercalating interaction mode. Conclusion: Our findings highlight 5-nitro-thiophene-thiosemicarbazone derivatives, especially LNN-05, as a promising new class of compounds for further studies to provide new anticancer therapies.

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The Effect of Phase Transition Temperature on Therapeutic Efficacy of Liposomal Bortezomib

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200101150640 ◽

2020 ◽

Vol 20 (6) ◽

pp. 700-708

Author(s):

Mitra Korani ◽

Sara Nikoofal-Sahlabadi ◽

Amin R. Nikpoor ◽

Solmaz Ghaffari ◽

Hossein Attar ◽

...

Keyword(s):

Side Effects ◽

Cell Line ◽

Therapeutic Efficacy ◽

Drug Loading ◽

Particle Diameter ◽

In Vitro Cytotoxicity ◽

Liposomal Formulations ◽

Cytotoxicity Studies ◽

Anti Tumor Activity

Aims: Here, three liposomal formulations of DPPC/DPPG/Chol/DSPE-mPEG2000 (F1), DPPC/DPPG/Chol (F2) and HSPC/DPPG/Chol/DSPE-mPEG2000 (F3) encapsulating BTZ were prepared and characterized in terms of their size, surface charge, drug loading, and release profile. Mannitol was used as a trapping agent to entrap the BTZ inside the liposomal core. The cytotoxicity and anti-tumor activity of formulations were investigated in vitro and in vivo in mice bearing tumor. Background: Bortezomib (BTZ) is an FDA approved proteasome inhibitor for the treatment of mantle cell lymphoma and multiple myeloma. The low solubility of BTZ has been responsible for the several side effects and low therapeutic efficacy of the drug. Encapsulating BTZ in a nano drug delivery system; helps overcome such issues. Among NDDSs, liposomes are promising diagnostic and therapeutic delivery vehicles in cancer treatment. Objective: Evaluating anti-tumor activity of bortezomib liposomal formulations. Methods: Data prompted us to design and develop three different liposomal formulations of BTZ based on Tm parameter, which determines liposomal stiffness. DPPC (Tm 41°C) and HSPC (Tm 55°C) lipids were chosen as variables associated with liposome rigidity. In vitro cytotoxicity assay was then carried out for the three designed liposomal formulations on C26 and B16F0, which are the colon and melanoma cancer mouse-cell lines, respectively. NIH 3T3 mouse embryonic fibroblast cell line was also used as a normal cell line. The therapeutic efficacy of these formulations was further assessed in mice tumor models. Result: MBTZ were successfully encapsulated into all the three liposomal formulations with a high entrapment efficacy of 60, 64, and 84% for F1, F2, and F3, respectively. The findings showed that liposomes mean particle diameter ranged from 103.4 to 146.8nm. In vitro cytotoxicity studies showed that liposomal-BTZ formulations had higher IC50 value in comparison to free BTZ. F2-liposomes with DPPC, having lower Tm of 41°C, showed much higher anti-tumor efficacy in mice models of C26 and B16F0 tumors compared to F3-HSPC liposomes with a Tm of 55°C. F2 formulation also enhanced mice survival compared with untreated groups, either in BALB/c or in C57BL/6 mice. Conclusion: Our findings indicated that F2-DPPC-liposomal formulations prepared with Tm close to body temperature seem to be effective in reducing the side effects and increasing the therapeutic efficacy of BTZ and merits further investigation.

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Characterization of a newly established uterine carcinosarcoma cell line featuring the sarcomatous phenotype of the tumor in vitro

International Journal of Gynecological Cancer ◽

10.1111/j.1525-1438.2007.01004.x ◽

2008 ◽

Vol 18 (2) ◽

pp. 339-344 ◽

Cited By ~ 6

Author(s):

H.-J. Schulten ◽

J. Wolf-Salgó ◽

C. Gründker ◽

B. Gunawan ◽

L. FÜZESI

Keyword(s):

Cell Line ◽

Hormone Receptors ◽

Tumor Biology ◽

Growth Factor Receptor ◽

Population Doubling ◽

Established Cell Line ◽

Population Doubling Time ◽

Uterine Carcinosarcoma ◽

Specific Staining

We describe the newly established cell line CS-99 derived from a uterine carcinosarcoma retaining features of the sarcomatous phenotype in vitro. CS-99 cells exhibit a mesenchymal morphology with predominantly spindle-shaped cells at nonconfluence turning to pleomorphic appearance at confluence. The mesenchymal phenotype was evidenced immunohistochemically by strong vimentin and moderate SM-actin, which was similar to the sarcomatous component of the primary tumor. P53 was overexpressed in a subset of CS-99 cells. Epithelial membrane antigen was moderately expressed whereas other markers including pan CK, CK 5/6, CK 34, epidermal growth factor receptor, desmin, carcinoembryonic antigen, S100, KIT, ERBB2, and the hormone receptors, estrogen receptor and progesterone receptor revealed either weak or no specific staining in CS-99 cells. High self-renewal capacity corresponded to the population doubling time of 23 h in high passage. CS-99 cells were able to develop three-dimensional tumor spheroids in vitro. Cytogenetic analysis and multicolor fluorescence in situ hybridization of CS-99 demonstrated an almost stable karyotype including numerical changes +8, +18, and +20 and translocations, amongst others der(1)t(1;2), der(1)t(1;7), der(2)t(2;19), der(5)t(5;8), and der(5)t(5;14). Taken together, the cell line CS-99 exhibits strong growths dynamics and a complex but stable karyotype in higher passages, and can be further a useful in vitro model system for studying tumor biology of carcinosarcomas.

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Antifungal Prenylated Diphenyl Ethers from Arthrinium arundinis, an Endophytic Fungus Isolated from the Leaves of Tobacco (Nicotiana tabacum L.)

Molecules ◽

10.3390/molecules23123179 ◽

2018 ◽

Vol 23 (12) ◽

pp. 3179 ◽

Cited By ~ 4

Author(s):

Peng Zhang ◽

Xin Li ◽

Xiao-Long Yuan ◽

Yong-Mei Du ◽

Bin-Gui Wang ◽

...

Keyword(s):

Nicotiana Tabacum ◽

Cell Line ◽

Endophytic Fungus ◽

In Vitro Cytotoxicity ◽

Ionization Mass ◽

Monocytic Cell ◽

Monocytic Cell Line ◽

Nicotiana Tabacum L ◽

Diphenyl Ethers

An endophytic fungus Arthrinium arundinis TE-3 was isolated and purified from the fresh leaves of cultivated tobacco (Nicotiana tabacum L.). Chemical investigation on this fungal strain afforded three new prenylated diphenyl ethers (1−3) as well as three known analogues (4−6). Structure elucidation of the isolated compounds was carried out by analysis of 1D and 2D nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectroscopy (HRESIMS) spectra, as well as by comparison of those data with literature data. The absolute configuration of the stereogenic center at C-8 in 1 was assigned by comparison of the experimental and calculated ECD spectra. Compounds 1 and 2 showed selective antifungal activity against Mucor hiemalis with minimum inhibitory concentration (MIC) values of 8 and 4 μg/mL, respectively. Compounds 5 and 6 exhibited inhibitory activity against Alteraria alternata with an MIC value of 8 μg/mL. In the cytotoxic assay, 2, 5, and 6 displayed moderate in vitro cytotoxicity against the human monocytic cell line (THP-1 cell line), with IC50 values of 40.2, 28.3, and 25.9 μM, respectively. This study indicated that endophytic fungi possess great potential for exploring new bioactive secondary metabolites.

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IN VITRO CYTOTOXICITY POTENTIAL OF ETHANOL EXTRACT OF SYZYGIUM SAMARANGENSE (Wt.)

Kongunadu Research Journal ◽

10.26524/krj283 ◽

2019 ◽

Vol 6 (1) ◽

pp. 30-32

Author(s):

Poonkodi K ◽

Mini R ◽

Vimaladevi K ◽

Prabhu V ◽

Anusuya M ◽

...

Keyword(s):

Fatty Acids ◽

Hela Cell ◽

Cell Line ◽

Ethanol Extract ◽

In Vitro Cytotoxicity ◽

Hela Cell Line ◽

Phytochemical Screening ◽

Ic50 Value ◽

Dependent Activity

The present investigation is carried out to study the invitro cytotoxicity of ethanol extract of Syzygium samarangense leaves on HeLa cell line by using MTT assay. Ethanol extract of S. samarangense showed concentration dependent activity on HeLa cell line with IC50 value of 40.5 μg/ml which shows that ethanol extract of S. samarangense posses significant cytoxicity.Moreover the preliminary phytochemical screening showed the presence of fatty acids, alkaloids, flavonoids, terphenoids, saponins, tannins and steroids which are responsible for its cytotoxicity. There are only a few reports are available for cytotoxicity of ethanol extract of S. samarangense.

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A COMPARATIVE ANTICANCER STUDY OF BIOSYNTHESIZED NANOPARTICLES, DOXORUBICIN & NANO-DOX CONJUGATE AGAINST CERVICAL CANCER HELA CELL LINE

10.36106/ijsr/2509427 ◽

2020 ◽

pp. 4-7

Author(s):

M. R. Kamala Priya ◽

Priya R. Iyer

Keyword(s):

Cervical Cancer ◽

Gold Nanoparticles ◽

Cell Death ◽

Cancer Therapy ◽

Hela Cell ◽

Cell Line ◽

In Vitro Cytotoxicity ◽

Hela Cell Line ◽

Vero Cell Line

Doxorubicin is the most common chemotherapy drug used in cancer therapy. Its usage is associated with various side-effects. In order to overcome the challenges in Doxorubicin administration, the present study has focussed on synthesizing a drug conjugate with biosynthesized gold nanoparticles and doxorubicin. The gold nanoparticles were biosynthesized using green extracts of medicinal plants with potential anticancer activities. The nanoparticle that possesses anticancer activity was conjugated with the drug for a combinatorial effect of the nanoparticles and the drug. The in vitro cytotoxicity was checked in Vero cell line through MTT assay. The in vitro anti proliferative effects were screened against cervical cancer in HeLa cell line. Fluorescence activated cell sorting analysis was carried out to detect the difference between live and dead cell populations. The preliminary confirmation was carried out in UV-VIS spectrophotometer. The morphological characterization was carried out by SEM and stability by Zeta potential. The IC50 of the nanocompounds demonstrated anti-proliferative activity against cervical cancer similar to the chemotherapeutic drug, Doxorubicin; additionally in a much lesser concentration of the drug. The in vitro cytotoxicity exhibited high viability of cells in Vero cell line with minimum viability of 80% in all the tested concentrations. There was a synergistic effect of the nanoparticles along with the drug; thereby an enhanced therapeutic efficiency was achieved. FACS analysis showed 36% of cell death in Dox treated HeLa cells whereas 96% of cell death in Nano-Dox treated HeLa cells. Nano-Dox conjugate has demonstrated high anticancer effects than drug alone Doxorubicin. Further biosynthesized nanomaterials based drug formulation can be developed as a potential strategy in cancer therapy.

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Enhancement of pheomelanogenesis by L-dopa in the mouse melanocyte cell line, TM10, in vitro

Journal of Cell Science ◽

10.1242/jcs.87.4.507 ◽

1987 ◽

Vol 87 (4) ◽

pp. 507-512

Author(s):

C. Sato ◽

S. Ito ◽

T. Takeuchi

Keyword(s):

Cell Line ◽

Growth Medium ◽

Yellow Pigment ◽

Black Pigment ◽

Established Cell Line ◽

Treatment Results ◽

Common Precursor ◽

Established Cell ◽

The Common

Cells of TM10, an established cell line, are melanocytes that contain equal amounts of eumelanin (black pigment) and pheomelanin (yellow pigment). The content of pheomelanin drastically increased when the cells were cultured in growth medium containing 0.2mM-L-dopa (L-dihydroxyphenylalanine), which is the common precursor for both eumelanogenesis and pheomelanogenesis. After this treatment, the amount of pheomelanin was 3.7-fold greater than that of control in TM10, whereas the amount of eumelanin changed very little. In contrast, 5-S-cysteinyl-dopa, which is the specific precursor for pheomelanogenesis downstream of L-dopa, did not cause preferential increase in pheomelanogenesis. Ultrastructural observations also confirmed these results; in 0.2mM-L-dopa, an increase in the number of pheomelanosomes was observed in the cytoplasm of TM10 cells. Our results also suggest that the L-dopa treatment results in a decrease in tyrosinase activity per melanosome.

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