scholarly journals PC12 and THP-1 Cell Lines as Neuronal and Microglia Model in Neurobiological Research

2021 ◽  
Vol 11 (9) ◽  
pp. 3729
Author(s):  
Katarzyna Balon ◽  
Benita Wiatrak
Keyword(s):  
Cell Culture ◽  
Dna Damage ◽  
Cell Lines ◽  
Inflammatory Factor ◽  
Research Model ◽  
Differentiated Cells ◽  
Undifferentiated Cells ◽  
Primary Cell Lines ◽  
Neurobiological Research ◽  
The Impact

Models based on cell cultures have become a useful tool in modern scientific research. Since primary cell lines are difficult to obtain and handle, neoplasm-derived lines like PC12 and THP-1 offer a cheap and flexible solution for neurobiological studies but require prior differentiation to serve as a neuronal or microglia model. PC12 cells constitute a suitable research model only after differentiation by incubation with nerve growth factor (NGF) and THP-1 cells after administering a differentiation factor such as phorbol 12-myristate-13-acetate (PMA). Still, quite often, studies are performed on these cancer cells without differentiation. The study aimed to assess the impact of PC12 or THP-1 cell differentiation on sensitivity to harmful factors such as Aβ25-35 (0.001–5 µM) (considered as one of the major detrimental factors in the pathophysiology of Alzheimer’s disease) or lipopolysaccharide (1–100 µM) (LPS; a pro-inflammatory factor of bacterial origin). Results showed that in most of the tests performed, the response of PC12 and THP-1 cells induced to differentiation varied significantly from the effect in undifferentiated cells. In general, differentiated cells showed greater sensitivity to harmful factors in terms of metabolic activity and DNA damage, while in the case of the free radicals, the results were heterogeneous. Obtained data emphasize the importance of proper differentiation of cell lines of neoplastic origin in neurobiological research and standardization of cell culture handling protocols to ensure reliable results.

scholarly journals TMOD-01. CHARACTERIZATION OF PATIENT-DERIVED PRIMARY CELL LINES AND XENOGRAFTS FOR GLIOBLASTOMA

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi262-vi262 ◽  
Author(s):  
Noriyuki Kijima ◽  
Daisuke Kanematsu ◽  
Tomoko Shofuda ◽  
Masahiro Nonaka ◽  
Ryoichi Iwata ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Surface ◽  
Cell Lines ◽  
Primary Cell Culture ◽  
Surface Marker ◽  
Primary Cell ◽  
Cell Surface Marker ◽  
High Expression ◽  
Glioblastoma Patient ◽  
Primary Cell Lines

Abstract Patient-derived primary cell culture and xenograft are essential tools for translational research for glioblastoma. However, characteristics of each patient derived cell line and xenograft is not extensively studied. In this study, we aim to analyze the characteristics of our glioblastoma patient-derived cell lines and xenografts based on cell surface markers and their differentiation patterns. We have established 20 glioblastoma primary cell culture lines by serum free medium containing EGF and bFGF and found that primary cell culture lines could be classified based on the expression of CD133 and CD44. Four cell lines had high expression of both CD133 and CD44. Eleven cell lines had high expression of only CD44, three cell lines had high expression of only CD133, two cell lines had low expression of both CD133 and CD44. In addition when we induce differentiation, these cell lines showed differentiation to both glial and neuronal differentiation, but differentiation patterns were different depending on each cell line. Four cell lines showed predominant neuronal differentiation and others showed predominant glial differentiation. We next investigated in vivo characteristics of glioblastoma patient derived xenografts from these established cell lines. We have injected these cell lines into NOD/Shi-scid IL2Rγ KO mouse and histopathologically analyzed characteristics of xenografts. Each xenograft well recapitulated histological features of original patients’ tumors and tumor cells remarkably invade through subventricular zone. These results suggest that glioblastoma patient derived primary cell lines and xenografts have different characteristics of cell surface marker expressions and differentiation patterns, thus can classify these cell lines depending on cell surface marker expressions and differentiation patterns. Further analysis is needed to examine the biological importance of the differences in cell surface marker expressions and differentiation patterns.

scholarly journals Development of Primary Cell Lines from Gill, Kidney, Spleen and Caudal Fin of Common Carp (Cyprinus carpio)

2020 ◽  
Vol 12 (1) ◽  
pp. 73
Author(s):  
Insariani Insariani ◽  
Trisniaty Trisniaty ◽  
Freddy Riatmono ◽  
Abdul Ghani
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Common Carp ◽  
Cyprinus Carpio ◽  
Primary Cell ◽  
Fish Cell ◽  
Caudal Fin ◽  
Common Carp Cyprinus Carpio ◽  
Carp Cyprinus Carpio ◽  
Primary Cell Lines

HighlihgtDevelop primary cultures derived From  tissue tails fins, gills, kidney and spleen from local Indonesian carp (Cyprinus carpio).Primary culture cell with L15  Mediacell cultures  consist of two type Fibroblast-like and epithelial –like cell AbstractThe fish cell lines technology have been developed for the interests of the fisheries world. This study aimed at developing a primary cell line from gill, kidney, spleen, and caudal fin of a common carp (Cyprinus carpio). A healthy common carp weighing 20 g (~1 month) was collected from the Cijeruk Fish Seed Center, Bogor. The development of primary cell lines from the gill, fin, tail, kidney and spleen tissue was performed in cell culture medium Leibovitz’s L-15 supplemented with 20% serum fetal bovine, 250 IU Penicillin, 250 µg / ml kanamycin sulfate and 2Mm L-Glutamine, and incubated at 28°C. Primary cell lines of caudal fin and gill began to form a monolayer on day 17 after culture. While the development of cell lines from kidney and spleen, although the initiation of cells and cells spread on the surface into a monolayer, was not perfect; therefore, the passage was unable to be done. Microscopic observations and Giemsa staining showed primary cell lines of caudal fin and gill based on cell morphology consisted of two cell types, fibroblast-like cells and epithelial-like cells. The first passage was done on day 17 when the confluence was more than 50%. The next passage was carried out every 3 weeks when confluence reached 70% -80%. The primary cell culture of gill was successfully passaged as much as 72 and the caudal fin was successfully passed as much as 89 times over 7 years. These new cell lines can be further used to propagate fish viruses and other biotechnology assays.

scholarly journals Differentiated Paju cells have increased resistance to toxic effects of potassium ionophores.

2004 ◽  
Vol 51 (2) ◽  
pp. 539-544 ◽  
Author(s):  
Vera Teplova ◽  
Elina Jääskeläinen ◽  
Mirja Salkinoja-Salonen ◽  
Nils-Erik L Saris ◽  
Martina Serlachius ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane ◽  
Tumor Cell Line ◽  
Differentiated Cells ◽  
Apoptotic Protein ◽  
Undifferentiated Cells ◽  
Elevated Expression ◽  
The Impact ◽  
Microbial Toxins

In this study we have investigated the impact of differentiation of neuronal cells on their sensitivity to microbial toxins. We used the human neural crest-derived tumor cell line Paju, which can be induced to differentiation in vitro by treatment with phorbol 12-myristate 13-acetate. Addition of the highly toxic potassium ionophores cereulide (4.5 and 9.0 ng/ml) or valinomycin (20 ng/ml), to cultures of undifferentiated Paju cells caused collapse of the mitochondrial membrane potential - measured with the fluorescent probe 5,5',6,6'-tetrachloro-1,1',3,3'-tetrabenzimidazole carbocyanine iodide (JC-1) followed by detachment of the cells and their apoptotic death. After induced differentiation of the Paju cells, their mitochondria retained the membrane potential upon exposure to the toxins and the cells displayed increased resistance to apoptosis as compared with undifferentiated cells. This effect may be caused by an elevated expression of the anti-apoptotic protein Bcl-2 and of the neuroprotective factor, stanniocalcin, in differentiated cells.

scholarly journals Establishment of B-Cell Lines Latently Infected with Reactivation-Competent Murine Gammaherpesvirus 68 Provides Evidence for Viral Alteration of a DNA Damage-Signaling Cascade

2008 ◽  
Vol 82 (15) ◽  
pp. 7688-7699 ◽  
Author(s):  
J. Craig Forrest ◽  
Samuel H. Speck
Keyword(s):  
Cell Culture ◽  
Dna Damage ◽  
B Cells ◽  
B Cell ◽  
Cell Lines ◽  
Molecular Mechanisms ◽  
Cells In Culture ◽  
Host Determinants ◽  
Dna Damage Signaling ◽  
Gammaherpesvirus 68

ABSTRACT Gammaherpesvirus 68 (γHV68, or MHV68) is a naturally occurring rodent pathogen that replicates to high titer in cell culture and is amenable to in vivo experimental evaluation of viral and host determinants of gammaherpesvirus disease. However, the inability of MHV68 to transform primary murine B cells in culture, the absence of a robust cell culture latency system, and the paucity of MHV68-positive tumor cell lines have limited an understanding of the molecular mechanisms by which MHV68 modulates the host cell during latency and reactivation. To facilitate a more complete understanding of viral and host determinants that regulate MHV68 latency and reactivation in B cells, we generated a recombinant MHV68 virus that encodes a hygromycin resistance protein fused to enhanced green fluorescent protein as a means to select cells in culture that harbor latent virus. We utilized this virus to infect the A20 murine mature B-cell line and evaluate reactivation competence following treatment with diverse stimuli to reveal viral gene expression, DNA replication, and production of progeny virions. Comparative analyses of parental and infected A20 cells indicated a correlation between infection and alterations in DNA damage signaling following etoposide treatment. The data described in this study highlight the potential utility of this new cell culture-based system to dissect molecular mechanisms that regulate MHV68 latency and reactivation, as well as having the potential of illuminating biochemical alterations that contribute to gammaherpesvirus pathogenesis. In addition, such cell lines may be of value in evaluating targeted therapies to gammaherpesvirus-related tumors.

scholarly journals CBMT-16. EGFRvIII EXPRESSION CONFERS CHEMOSENSITIVITY BY INCREASING DNA MISMATCH REPAIR PROTEIN EXPRESSION AND REPLICATION STRESS

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi36-vi36
Author(s):  
Nina Struve ◽  
Zev A Binder ◽  
Lucy F Stead ◽  
Tim Brend ◽  
Stephen J Bagley ◽  
...  
Keyword(s):  
Dna Damage ◽  
Protein Expression ◽  
Treatment Response ◽  
Mismatch Repair ◽  
Cell Lines ◽  
Dna Mismatch Repair ◽  
Replication Stress ◽  
Dna Mismatch ◽  
Mgmt Promoter ◽  
The Impact

Abstract MGMT promoter methylation is the only accepted biomarker with prognostic role in GBM but its routine implementation is limited partly response to TMZ is heterogeneous, but also due to lack of effective alternative treatment options. Therefore, additional biomarkers are needed to enable better prediction of survival and to improve individualized treatment of GBM patients. A potential new biomarker is the epidermal growth factor receptor variant III (EGFRvIII). This constitutively activated deletion variant is present in approximately one third of all IDH wildtype GBM, but its relevance to treatment response is poorly understood. The aim of the present study was to analyze the impact of endogenous EGFRvIII expression on chemosensitivity and the mechanisms underlying any differential treatment response. EGFRvIII expression was associated with prolonged median overall survival but only for GBM patients with MGMT promoter methylated tumors. In line with this, we observed increased TMZ sensitivity of EGFRvIII+ and MGMT promoter methylated cells, which translated into improved survival in xenograft experiments. The increased TMZ sensitivity was associated with an elevated DNA damage induction accompanied by an increased expression of DNA mismatch repair (MMR) proteins in EGFRvIII+ cell lines and EGFRvIII+ GBM patient samples. Subsequently, only a moderate reduction in MMR protein expression resulted in a dramatic TMZ resistance, suggesting that EGFRvIII expression specifically sensitized MGMT deficient cells to TMZ treatment by upregulating MMR. Furthermore, EGFRvIII expression in GBM cell lines was accompanied by increased DNA damage, replication fork slowing, stalling and enhanced origin firing, implying replication stress. Targeting of EGFRvIII-dependent replication stress by irinotecan led to hypersensitivity of EGFRvIII+ cells. Taken together this study illustrates that EGFRvIII-induced upregulation of MMR and replication stress increases chemosensitivity thereby highlighting the vulnerability of EGFRvIII+ GBM to available treatments. These important data may also guide the development of new and more effective personalized strategies.

scholarly journals Co-occurrence of Cell Lines, Basal Media and Supplementation in the Biomedical Research Literature

2020 ◽  
Vol 5 (3) ◽  
pp. 161-177 ◽  
Author(s):  
Jessica Cox ◽  
Darin McBeath ◽  
Corey Harper ◽  
Ron Daniel
Keyword(s):  
Cell Culture ◽  
Text Mining ◽  
Cell Lines ◽  
Biomedical Research ◽  
Research Literature ◽  
Sentence Structure ◽  
In Vitro Cell Culture ◽  
Basal Media ◽  
The Impact

AbstractPurposeThe use of in vitro cell culture and experimentation is a cornerstone of biomedical research, however, more attention has recently been given to the potential consequences of using such artificial basal medias and undefined supplements. As a first step towards better understanding and measuring the impact these systems have on experimental results, we use text mining to capture typical research practices and trends around cell culture.Design/methodology/approachTo measure the scale of in vitro cell culture use, we have analyzed a corpus of 94,695 research articles that appear in biomedical research journals published in ScienceDirect from 2000–2018. Central to our investigation is the observation that studies using cell culture describe conditions using the typical sentence structure of cell line, basal media, and supplemented compounds. Here we tag our corpus with a curated list of basal medias and the Cellosaurus ontology using the Aho-Corasick algorithm. We also processed the corpus with Stanford CoreNLP to find nouns that follow the basal media, in an attempt to identify supplements used.FindingsInterestingly, we find that researchers frequently use DMEM even if a cell line's vendor recommends less concentrated media. We see long-tailed distributions for the usage of media and cell lines, with DMEM and RPMI dominating the media, and HEK293, HEK293T, and HeLa dominating cell lines used.Research limitationsOur analysis was restricted to documents in ScienceDirect, and our text mining method achieved high recall but low precision and mandated manual inspection of many tokens.Practical implicationsOur findings document current cell culture practices in the biomedical research community, which can be used as a resource for future experimental design.Originality/valueNo other work has taken a text mining approach to surveying cell culture practices in biomedical research.

scholarly journals Defining the mechanistic toxicity of metal oxide nanoparticles in vitro, under physiologically relevant oxygen concentrations

2021 ◽  
Author(s):  
◽  
Michael Theodoulides
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Hepg2 Cells ◽  
Metal Oxide Nanoparticles ◽  
Culture Conditions ◽  
Oxide Nanoparticles ◽  
Cell Type ◽  
Culture Environment ◽  
The Impact ◽  
Cell Culture Conditions

Metal oxide nanoparticles (MONPs) are intended for use in numerous consumer applications, leading to inevitable human exposure. Previous work conducted in hyperoxic cell culture conditions (21% O2, 5% CO2) with nanoparticles (NPs) has proven the ability of some material types to induce genotoxicity and inflammatoxicity. Alteration in intracellular calcium [i(Ca2+)] signalling is involved in facilitating toxicity through the alteration of signal-transduction pathways, but there is less understanding of the impact of NPs exposure upon changes in such signalling pathways. Furthermore, whilst human cells cultured in ambient air may induce a particular toxicity profile, this may not be the same under the physiologic oxygen conditions experienced in the human body. Therefore, the aim of this study was to assess the impact of anatase TiO2 (NM-102), Rutile TiO2 (NM-104) and dextran coated superparamagnetic Fe3O4 (dSPIONs) upon monocytes (THP-1), macrophages (dTHP-1) and hepatocarcinoma (HepG2) cells in both an in vivo-resembling physioxia environment (5%O2, 5%CO2) and hyperoxic cell culture conditions (21%O2, 5%CO2). Their impact on i(Ca2+) homeostasis and how it relates to their potential genotoxic potential was also evaluated.Due to the importance of different physicochemical characteristics for the facilitation of toxicity, all MONPs were characterized. MONPs hydrodynamic diameter (HD) and ζ-potential (ζ) in PBS were identified using dynamic light scattering: NM-102: HD=391.9nm, ζ =7.1±2.0mV; NM-104: HD=255nm, ζ=14.6 +/- 2.1mV; dSPIONs: HD=88.6nm, ζ =10.4±1.3mV. The possible toxic effect of NPs depends on their concentration and duration of their interaction with cells. Therefore, following 24h exposure to dSPIONs (0-100µg/ml), concentration-dependent and cell-type-dependent (dTHP1>THP-1>HepG2) significant increases in NP-cellular interaction were observed, which was significantly greater in the physioxic culture environment. Concurrent, significant loss of dSPION-associated cell proliferation (evaluated using relative population doubling) in all cell lines and significant increases in DNA damage was also identified in HepG2 cells (using the cytokinesis block micronucleus assay), albeit only in physioxia. Exposure to ≥10ug/ml NM-102 and NM-104 resulted in significant, two-fold increases in micronuclei formation in HepG2 in both environments. All MONPs induced a significant increase in tumour necrosis factor-α and interleukin-8 secretion in all cell lines and oxygen culture environments. Increase in the production of the chemokines was correlated with the observed HepG2 cell genotoxicity. In all cell lines and cell culture environments, treatment for up to 5h with NM-102 or dSPIONs triggered cell type specific increases in i(Ca2+) that correlated with the reduction of cellular antioxidant glutathione (measured after 5h treatment with all the MONPs). After pre-treatment of the cell lines with antioxidant trollox in all cell culture environments i(Ca2+) appeared to be increased independently from the change of cellular redox status. Environment-specific biological interaction and impacts with regard to NP uptake, genotoxic effects, and consequence on cellular signaling mechanisms were only observed with dSPIONs in a physioxic culture environment, while NM-102 and NM-104 induced similar effects in both environments. The results presented in this study allow the conclusion that the environmental oxygen content has an impact on the NP toxicity profiles although it is NP dependent.

Preclinical evidence of ET-743 as a potential chemotherapy option for the treatment of biliary carcinoma.

2013 ◽  
Vol 31 (4_suppl) ◽  
pp. 193-193
Author(s):  
Francesco Leone ◽  
Caterina Peraldo-Neia ◽  
Giuliana Cavalloni ◽  
Marco Soster ◽  
Loretta Gammaitoni ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Culture ◽  
Dna Damage ◽  
Clinical Trials ◽  
Cell Lines ◽  
Primary Cell Culture ◽  
Primary Cell ◽  
Term Survival ◽  
Anti Tumor Activity

193 Background: The standard chemotherapy for unresectablebiliary tract carcinoma (BTC) is based on gemcitabine and platinum compounds. However, these combinations have not been shown to be effective in improving long-term survival. Thus, there is a real need to find new strategies that would impact in a significant way on clinical outcome. Ecteinascidin-743 (ET-743), a compound isolated from the marine tunicate Ecteinascidia turbinata. ET-743, is approved for the treatment of ovarian cancer and soft tissue sarcoma. Phase II and III clinical trials are ongoing for the treatment of different solid tumors. No preclinical data are available about the efficacy of ET-743 in BTC. In a phase I study, one patient received ET-743 plus capecitabine and experienced a long lasting complete metabolic response. Here, we investigated the antitumor activity of ET-743 in preclinical BTC models. Methods: Four BTC cell lines TFK1, EGI-1, HuH28 and TGBC1 were used to evaluate the effect of ET-743 on proliferation, cell cycle, apoptosis and on the activation of DNA damage proteins. The effect on proliferation was also investigated on a primary cell culture of a gallbladder carcinoma (GBC) resistant to gemcitabine and oxaliplatin. On the same cells, the inhibition of VEGF secretion mediated by ET-743 was analyzed by ELISA. The anti-tumor activity of ET-743 was tested on EGI-1 xenografts in NOD/SCID mice. Results: In vitro, ET-743 is able to markedly reduce cell proliferation of BTC cell lines through cell cycle blockage on G0/G1 phase and to inhibit the growth of primary cell culture derived from GBC patient. Moreover, ET-743 promotes apoptosis by caspase 3 activation, activates proteins involved in DNA damage and reduces VEGF secretion. In the in vivo model, ET-743 is able to slow tumor growth in BTC xenograft. The mechanism of anti-tumor activity involves DNA damage, the induction of hypoxia transcription factor-1, and angiogenesis inhibition. ET-743 has no significant effect on apoptosis in vivo. Conclusions: These data suggest that ET-743 could represent an alternative chemotherapy for BTC treatment and encourage the development of clinical trials of ET-743 in BTC patients.

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