scholarly journals Assessment of Intratumoral Heterogeneity in Isolated Human Primary High-Grade Glioma: Cluster of Differentiation 133 and Cluster of Differentiation 15 Double Staining of Glioblastoma Subpopulations

2021 ◽  
Vol 9 (A) ◽  
pp. 87-94
Author(s):  
Ahmad Faried ◽  
Wahyu Widowati ◽  
Rizal Rizal ◽  
Hendrikus M. B. Bolly ◽  
Danny Halim ◽  
...  
Keyword(s):  
Brain Tumors ◽  
Cell Line ◽  
Cell Lines ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Double Staining ◽  
Glioblastoma Cells ◽  
Ki 67 ◽  
Primary Glioblastoma ◽  
Cluster Of Differentiation

BACKGROUND: Gliomas are the most common primary brain tumors, representing 50–60% of malignant primary brain tumors. Gliomas are highly heterogeneous with marked inter- and intratumoral diversity. Gliomas heterogeneity is a challenging issue in the development of personalized treatment. The simplest method for studying heterogeneity is using ex vivo cell cultures; in our case, the cell lines were isolated from patient with glioblastomas. AIM: Here, we reported distinct cell subpopulations heterogeneity in glioblastoma cells. METHODS: Human glioblastoma cells isolation is conducted by enzymatic method with combination of collagenase I, hyaluronidase, and trypsin enzyme in proportional amount from patient. Immunostaining was performed to assess glial fibrillary acidic protein (GFAP), Ki-67, isocitrate dehydrogenase-1 (IDH-1) status, and program death ligand-1 (PD-L1) expression. Primary glioblastoma cell line was characterized by flow cytometry (fluorescence-activated cell sorting) analysis based on cluster of differentiation (CD) 133 and CD15 marker expression. U87MG and CGNH-89 cell lines were used as control. Distinct subpopulation analysis was performed by double staining of CD133 and CD15 in isolated primary glioblastoma cell line and its comparative control cells. RESULTS: Our isolated glioblastoma cells morphology was adherent cells which were able to form spheres depending on environment. Immunostaining confirmed GFAP, Ki-67, IDH-1 mutants, and PD-L1 expression. Our isolated glioblastoma cells expressed CD133 and CD15, coexpressed CD133/CD15 in different patterns. The highest subpopulation in primary glioblastoma was CD133+/CD15+. CONCLUSION: Glioblastoma cells can be isolated using enzymatic methods. Isolated glioblastoma cells consist of four different subpopulations distinguished by CD133/CD15 double staining. Intratumoral heterogeneity exists and directly or indirectly depends on their microenvironment.

scholarly journals Circumdatin-Aspyrone Conjugates from the Coral-Associated Aspergillus ochraceus LCJ11-102

Marine Drugs ◽  
2019 ◽  
Vol 17 (7) ◽  
pp. 400 ◽  
Author(s):  
Yaqin Fan ◽  
Yalin Zhou ◽  
Yuqi Du ◽  
Yi Wang ◽  
Peng Fu ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Nucleophilic Addition ◽  
Human Cancer ◽  
Aspergillus Ochraceus ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Human Cancer Cell ◽  
Human Cancer Cell Lines

Ochrazepines A−D (1−4), four new conjugates dimerized from 2-hydroxycircumdatin C (5) and aspyrone (6) by a nucleophilic addition to epoxide, were isolated from the fermentation broth of the coral-associated Aspergillus ochraceus strain LCJ11-102. Their structures including absolute configurations were determined based on spectroscopic analysis and chemical methods. Compounds 1−4 were also obtained by the semisynthesis from a nucleophilic addition of 2-hydroxycircumdatin C (5) to aspyrone (6). New compound 1 exhibited cytotoxic activity against 10 human cancer cell lines while new compounds 2 and 4 selectively inhibited U251 (human glioblastoma cell line) and compound 3 was active against A673 (human rhabdomyoma cell line), U87 (human glioblastoma cell line), and Hep3B (human liver cancer cell line) with IC50 (half maximal inhibitory concentration) values of 2.5–11.3 μM among 26 tested human cancer cell lines.

P13.09 Inconsistent effect of temozolomide exposure on cell viability in glioblastoma cell line models - a systematic review

2021 ◽  
Vol 23 (Supplement_2) ◽  
pp. ii34-ii34
Author(s):  
M M C Bruce ◽  
M T C Poon ◽  
P M Brennan
Keyword(s):  
Systematic Review ◽  
Cell Viability ◽  
Cell Line ◽  
Cell Lines ◽  
Exposure Duration ◽  
Laboratory Research ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Glioblastoma Cell Lines ◽  
Common Cell

Abstract BACKGROUND The alkylating agent temozolomide is part of standard care for patients with glioblastoma. Potential novel therapeutic agents are often first evaluated against temozolomide in glioblastoma cell line models. Despite the importance of this step in compound development, there is no standard concentration or exposure duration of temozolomide in laboratory research, and consistency in the effect of temozolomide on glioblastoma cell lines has not been assessed. This systematic review aimed to summarise the concentration and exposure duration of temozolomide and its effect on cell viability in studies using glioblastoma cell lines. MATERIAL AND METHODS We searched Medline and Embase Jan 1994 - Feb 2021 for studies that used at least one glioblastoma cell line and reported a measure of cell viability associated with temozolomide exposure. Studies were excluded if they used modified cell lines or did not report a cell viability measure associated with temozolomide as monotherapy. One reviewer screened all records and two reviewers assessed potentially eligible studies for inclusion. The main data items included the cell lines used, the concentration and exposure duration to temozolomide, and cell viability measures. We summarised findings using descriptive statistics. RESULTS Of 1,533 potentially eligible studies we included 213 studies reporting 209 different cell lines. The most common cell lines were U87, U251 and T98G, used in 61%, 41%, and 27% of studies, respectively. Twenty-five (12%) studies used patient-derived cell lines. The concentration of temozolomide used ranged from 0 to 8000μM. The temozolomide exposure duration ranged from <24 hours to >96 hours, with 29% studies using 72 hours. The most common cell viability measure was half maximal inhibitory concentration (IC50), which was reported in 183 (86%) studies. The median IC50 in 32 studies using the U87 cell line was 180μM (interquartile range [IQR]: 52–254μM) at 48-hour temozolomide exposure and 202μM (IQR 52–518μM) at 72-hour exposure. The median IC50 in 31 studies using U251 cell line was 84μM (IQR: 34–324μM) at 48-hour exposure and 102μM (IQR: 35–358μM) at 72-hour exposure. CONCLUSION Experimental setup of temozolomide and its effect on cell viability vary widely between studies using similar glioblastoma cell lines. This inconsistency of response to temozolomide questions reproducibility and the translational value of study findings.

Anticancer Potential and Mechanism of Action of Mango Ginger (Curcuma amada Roxb.) Supercritical CO2 Extract in Human Glioblastoma Cells

2014 ◽  
Vol 20 (2) ◽  
pp. 109-119 ◽  
Author(s):  
Cheppail Ramachandran ◽  
Ivonne V. Lollett ◽  
Enrique Escalon ◽  
Karl-Werner Quirin ◽  
Steven J. Melnick
Keyword(s):  
Cell Line ◽  
Mechanism Of Action ◽  
Dependent Manner ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Glioblastoma Cells ◽  
Embryonic Mouse ◽  
Human Glioblastoma ◽  
Mango Ginger ◽  
Curcuma Amada

Mango ginger ( Curcuma amada Roxb.) is among the less-investigated species of Curcuma for anticancer properties. We have investigated the anticancer potential and the mechanism of action of a supercritical CO2 extract of mango ginger (CA) in the U-87MG human glioblastoma cell line. CA demonstrated higher cytotoxicity than temozolomide, etoposide, curcumin, and turmeric force with IC50, IC75, and IC90 values of 4.92 μg/mL, 12.87 μg/mL, and 21.30 μg/mL, respectively. Inhibitory concentration values of CA for normal embryonic mouse hypothalamus cell line (mHypoE-N1) is significantly higher than glioblastoma cell line, indicating the specificity of CA against brain tumor cells. CompuSyn analysis indicates that CA acts synergistically with temozolomide and etoposide for the cytotoxicity with combination index values of <1. CA treatment also induces apoptosis in glioblastoma cells in a dose-dependent manner and downregulates genes associated with apoptosis, cell proliferation, telomerase activity, oncogenesis, and drug resistance in glioblastoma cells.

scholarly journals Multiple Localization of Endogenous MARK4L Protein in Human Glioma

2009 ◽  
Vol 31 (5) ◽  
pp. 357-370
Author(s):  
Ivana Magnani ◽  
Chiara Novielli ◽  
Melissa Bellini ◽  
Gaia Roversi ◽  
Lorenzo Bello ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Cellular Localization ◽  
Protein Identification ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Human Glioma ◽  
Glioma Cell Lines ◽  
Biochemical Fractionation

Background: We have previously shown that the sustained expression of MARK4L transcripts in glioma and neural progenitors (NHNPs) declines after exposure to antisense MARK4L oligonucleotides in glioblastoma cell lines. Array-CGH confirmed the genomic duplication of MARK4L identified by FISH in a glioblastoma cell line. This background together with literature data on the exogenous association of MARK4 with interphase centrosome prompted us to investigate the sub-cellular localization of the endogenous MARK4L protein aiming at achieving insights on its possible role in the pathomechanisms of glioma.Methods: Immunodetection was carried out to validate the specificity of MARK4L antibody in gliomas and NHNPs. Mass spectrometry was applied for MARK4L protein identification in a representative glioblastoma cell line. Combined biochemical fractionation and immunodetection analyses were performed to confirm the sub-cellular localization of MARK4L achieved by immunofluorescence in glioma cell lines.Results: By assigning MARK4L protein within the band immunoprecipitated by the specific antibody we validated our anti-MARK4L antibody. We demonstrated that the endogenous MARK4L: (i) colocalizes with centrosomes at all mitotic stages and resides in centrosome-enriched fractions; (ii) associates with the nucleolus and the midbody and respective fractions, and (iii) co-stains the aberrant centrosome configurations observed in glioma cell lines.Conclusions: The overall data merge on the multiplex entry of MARK4L into the cell cycle and link it to the aberrant centrosomes in glioma cell lines suggesting a possible role of this kinase in the abnormal mitotic processes of human glioma.

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 In-vitro Study: Camp Overshoot Caused by Chronic Morphine Alleviated by the Synergistic Combination of Zamzam Water and Methadone in Human Primary Glioblastoma Cell Line (U-87 MG)

2020 ◽  
Vol 18 (2) ◽  
Author(s):  
Shariff Halim ◽  
Nasir Mohamad ◽  
Ridzuan PM ◽  
Nur Husna Zakaria ◽  
Nur Atikah Muhammad
Keyword(s):  
Cell Line ◽  
Time Course ◽  
Camp Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Morphine Treatment ◽  
Chronic Morphine ◽  
Primary Glioblastoma ◽  
Chronic Morphine Treatment

Introduction: Zamzam water is blessed water originated from Mecca and is believed by Muslims to have the ability to cure illness. This ability to cure illness is due to the fact that zamzam water has higher concentration of minerals especially sodium, calcium and magnesium which play a vital role. Sodium is reported to be involved in the regulation of the Mu-Opioid Receptor, which indirectly leads to the production of cAMP. Hence, this current study was carried out to investigate the synergistic effects of zamzam water and methadone combination on cAMP levels in human primary glioblastoma cell line (U-87 MG) after chronic morphine administration. Materials and Methods: The time course and concentration of morphine on U-87 MG cell line was determined. The U-87 MG cell line was incubated with morphine (25 µL/mL) for 24h, to make the cell dependent on morphine and later treated with different combinations of 3.2 mL of zamzam water and methadone (5, 10, 25 µL/mL). The levels of cAMP was determined using the enzyme-linked immunosorbent assay kit. Results: The result revealed that 3.2 mL of zamzam water incubated with 10 µl/ mL of methadone significantly prevented the overshoot production of cAMP level (p<0.05) in U-87 MG cell line after 48h incubation when compared to the untreated samples. Conclusion: These finding suggest that co-treatment with zamzam water and methadone could possibly avoid tolerance and dependence on chronic morphine treatment by preventing the up-regulation of cAMP level.><0.05 in U-87 MG cell line after 48h incubation when compared to the untreated samples. Conclusion: These finding suggest that co-treatment with zamzam water and methadone could possibly avoid tolerance and dependence on chronic morphine treatment by preventing the up-regulation of cAMP level.

scholarly journals The suitability of glioblastoma cell lines as models for primary glioblastoma cell metabolism

2020 ◽  
Author(s):  
Anya Lara Arthurs ◽  
Damien J Keating ◽  
Simon J Conn
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Metabolic Flux ◽  
Krebs Cycle ◽  
Neural Cells ◽  
Glioblastoma Cell ◽  
Preclinical Research ◽  
Primary Glioblastoma

Abstract Background In contrast to most non-malignant cells, cells comprising Glioblastoma multiforme (GBM), a deadly brain tumour with extremely poor prognosis, preferentially utilise glycolysis over oxidative phosphorylation for metabolism in a phenomenon known as the ‘Warburg effect’. As effective treatments for GBM are severely lacking, research into therapeutics targeting the disease’s highly glycolytic state offer a promising avenue to improve patient survival. These studies often employ GBM cell lines for in vitro studies which translate poorly to the in vivo patient context. Methods The metabolic traits of the seven most commonly used GBM cell lines were assessed using a Seahorse Bioscience Metabolic Flux Analyser and compared to primary GBM cells and primary healthy mixed neural cells from the same patients. Results In support of the glycolytic nature of the patient-derived GBM cell lines, basal mitochondrial rate (p = 0.043) and ATP-linked respiration (p < 0.001) were significantly lower than primary adjacent normal cells from the same patient and reserve capacity (p = 0.037) and Krebs Cycle capacity (p = 0.002) were significantly higher for 12 patients. While no cell line was found to accurately replicate all metabolic attributes of primary GBM cells, specific parameters could be modelled by specific lines. Conclusions U251MG, U373MG and D54 lines are recommended for researching mitochondrial metabolism, and the D645 line for researching ATP-linked respiration. The T98G cell line recapitulated glycolysis-related metabolic parameters of the primary GBM cells and is recommended for research relating to glycolysis. These findings can guide preclinical research into the development of novel therapeutics targeting metabolic pathways in GBM.

Anticarcinogenic and metal chelation properties of novel hydroxybenzylidene-1-indanone derivatives in the U-251 glioblastoma cell line

2018 ◽  
Vol 42 (5) ◽  
pp. 3878-3884 ◽  
Author(s):  
Mariana Lozano-Gonzalez ◽  
María Teresa Ramírez-Apan ◽  
Antonio Nieto-Camacho ◽  
Ruben Alfredo Toscano ◽  
Ana Laura Sanchez-Sandoval ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Cytotoxic Agents ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Metal Chelation ◽  
Line P

A novel series of (Z)-2-(hydroxy(aryl)methylene)-2,3-dihydro-1H-indanone derivatives were designed, synthesized and evaluated as cytotoxic agents against six cancer cell lines.

scholarly journals In Vitro/In Silico Study on the Role of Doubling Time Heterogeneity among Primary Glioblastoma Cell Lines

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
M.-E. Oraiopoulou ◽  
E. Tzamali ◽  
G. Tzedakis ◽  
A. Vakis ◽  
J. Papamatheakis ◽  
...  
Keyword(s):  
Cell Lines ◽  
In Silico ◽  
Intratumoral Heterogeneity ◽  
Patient Specific ◽  
Cell Physiology ◽  
Glioblastoma Cell ◽  
Glioblastoma Cells ◽  
Primary Glioblastoma ◽  
Glioblastoma Cell Lines

The application of accurate cancer predictive algorithms validated with experimental data is a field concerning both basic researchers and clinicians, especially regarding a highly aggressive form of cancer, such as Glioblastoma. In an aim to enhance prediction accuracy in realistic patient-specific environments, accounting for both inter- and intratumoral heterogeneity, we use patient-derived Glioblastoma cells from different patients. We focus on cell proliferation using in vitro experiments to estimate cell doubling times and sizes for established primary Glioblastoma cell lines. A preclinically driven mathematical model parametrization is accomplished by taking into account the experimental measurements. As a control cell line we use the well-studied U87MG cells. Both in vitro and in silico results presented support that the variance between tumor staging can be attributed to the differential proliferative capacity of the different Glioblastoma cells. More specifically, the intratumoral heterogeneity together with the overall proliferation reflected in both the proliferation rate and the mechanical cell contact inhibition can predict the in vitro evolution of different Glioblastoma cell lines growing under the same conditions. Undoubtedly, additional imaging techniques capable of providing spatial information of tumor cell physiology and microenvironment will enhance our understanding regarding Glioblastoma nature and verify and further improve our predictability.

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