Online Verification of Human Cell Line Identity by STR DNA Typing

Dirks, Wilhelm G.; Drexler, Hans

doi:10.1007/978-1-61779-080-5_5

Cited by 22 publications

(25 citation statements)

References 12 publications

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“…These tests function by confirming retention of short-nucleotide tandem repeats (STR) within the genomic DNA and are relatively rapid and highly confirmatory. 34 Changes in the STR results can indicate cell line contamination, changes in chromosome structure, or chromosome deletions, all of which can lead to variability in the ENM test result.…”

Section: Cause and Effect Diagram And Control Experiments For The Mtsmentioning

confidence: 99%

Use of Cause-and-Effect Analysis to Design a High-Quality Nanocytotoxicology Assay

Rösslein

Elliott

Salit

et al. 2015

Chem. Res. Toxicol.

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An important consideration in developing standards and regulations that govern the production and use of commercial nanoscale materials is the development of robust and reliable measurements to monitor the potential adverse biological effects of such products. These measurements typically require cell-based and other biological assays that provide an assessment of the risks associated with the nanomaterial of interest. In this perspective, we describe the use of cause-and-effect (C&E) analysis to design robust, high quality cell-based assays to test nanoparticle-related cytotoxicity. C&E analysis of an assay system identifies the sources of variability that influence the test result. These sources can then be used to design control experiments that aid in establishing the validity of a test result. We demonstrate the application of C&E analysis to the commonly used 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) cell-viability assay. This is the first time to our knowledge that C&E analysis has been used to characterize a cell-based toxicity assay. We propose the use of a 96-well plate layout which incorporates a range of control experiments to assess multiple factors such as nanomaterial interference, pipetting accuracy, cell seeding density, and instrument performance, and demonstrate the performance of the assay using the plate layout in a case study. While the plate layout was formulated specifically for the MTS assay, it is applicable to other cytotoxicity, ecotoxicity (i.e., bacteria toxicity), and nanotoxicity assays after assay-specific modifications.

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Section: Cause and Effect Diagram And Control Experiments For The Mtsmentioning

confidence: 99%

Use of Cause-and-Effect Analysis to Design a High-Quality Nanocytotoxicology Assay

Rösslein

Elliott

Salit

et al. 2015

Chem. Res. Toxicol.

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“…All human ovarian cancer cell lines were of epithelial origin. Cell lines were authenticated every half year by short tandem repeat (STR) DNA markers as described previously (22). In brief, PCR amplification of 8 highly polymorphic microsatellite STR loci and gender determination were measured, and the uniqueness of DNA profiles was compared for identity control within the STR database of ATCC (23).…”

Section: Tumor Cell Line Culturementioning

confidence: 99%

Chemotherapy Alters Monocyte Differentiation to Favor Generation of Cancer-Supporting M2 Macrophages in the Tumor Microenvironment

et al. 2013

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Current therapy of gynecologic malignancies consists of platinum-containing chemotherapy. Resistance to therapy is associated with increased levels of interleukin (IL)-6 and prostaglandin E2 (PGE 2 ), 2 inflammatory mediators known to skew differentiation of monocytes to tumor-promoting M2 macrophages. We investigated the impact of cisplatin and carboplatin on 10 different cervical and ovarian cancer cell lines as well as on the ability of the tumor cells to affect the differentiation and function of cocultured monocytes in vitro. Treatment with cisplatin or carboplatin increased the potency of tumor cell lines to induce IL-10-producing M2 macrophages, which displayed increased levels of activated STAT3 due to tumor-produced IL-6 as well as decreased levels of activated STAT1 and STAT6 related to the PGE 2 production of tumor cells. Blockade of canonical NF-kB signaling showed that the effect of the chemotherapy was abrogated, preventing the subsequent increased production of PGE 2 and/or IL-6 by the tumor cell lines. Treatment with the COX-inhibitor indomethacin and/or the clinical monoclonal antibody against interleukin-6 receptor (IL-6R), tocilizumab, prevented M2-differentiation. Importantly, no correlation existed between the production of PGE 2 or IL-6 by cancer cells and their resistance to chemotherapy-induced cell death, indicating that other mechanisms underlie the reported chemoresistance of tumors producing these factors. Our data suggest that a chemotherapy-mediated increase in tumor-promoting M2 macrophages may form an indirect mechanism for chemoresistance. Hence, concomitant therapy with COX inhibitors and/or IL-6R antibodies might increase the clinical effect of platinum-based chemotherapy in otherwise resistant tumors. Cancer Res; 73(8); 2480-92. Ó2013 AACR.

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“…The head and neck cancer cell lines UM-SCC1, UM-SCC25, UM-SCC6, UM-SCC11 that harbor wild-type HRAS, and PI3KCA were obtained from the University of Michigan (11). All cell lines were authenticated by DNA typing and used within 6 months of receipt (12). The mutation status of HRAS and PIK3CA genes was verified by sanger sequencing in all cell lines.…”

Section: Cell Lines Reagents and Mtt Viability Assaysmentioning

confidence: 99%

RAS/PI3K Crosstalk and Cetuximab Resistance in Head and Neck Squamous Cell Carcinoma

Ράμπιας

Giagini

Siolos

et al. 2014

Clinical Cancer Research

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Purpose: Cetuximab, an antibody directed against the EGF receptor, is an effective clinical therapy for patients with head and neck squamous cell cancer (HNSCC). Despite great clinical promise, intrinsic or acquired cetuximab resistance hinders successful treatment outcomes but little is known about the underlying mechanism.Experimental Design: To study the role of oncogenic HRAS in cetuximab resistance in HNSCC, the frequency of oncogenic HRAS mutations was determined in a cohort of 180 genomic DNAs from head and neck cancer specimens. We also used a combination of cetuximab-resistant cell lines and a transgenic mouse model of RAS-driven oral cancer to identify an oncogenic RAS-specific gene expression signature that promotes cetuximab resistance.Results: Here, we show that activation of RAS signaling leads to persistent extracellular signal-regulated kinase 1/2 signaling and consequently to cetuximab resistance. HRAS depletion in cells containing oncogenic HRAS or PIK3CA restored cetuximab sensitivity. In our study, the gene expression signature of c-MYC, BCL-2, BCL-XL, and cyclin D1 upon activation of MAPK signaling was not altered by cetuximab treatment, suggesting that this signature may have a pivotal role in cetuximab resistance of RAS-activated HNSCC. Finally, a subset of patients with head and neck cancer with oncogenic HRAS mutations was found to exhibit de novo resistance to cetuximab-based therapy.Conclusions: Collectively, these findings identify a distinct cetuximab resistance mechanism. Oncogenic HRAS in HNSCC promotes activation of ERK signaling, which in turn mediates cetuximab resistance through a specific gene expression signature.

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Online Verification of Human Cell Line Identity by STR DNA Typing

Cited by 22 publications

References 12 publications

Use of Cause-and-Effect Analysis to Design a High-Quality Nanocytotoxicology Assay

Use of Cause-and-Effect Analysis to Design a High-Quality Nanocytotoxicology Assay

Chemotherapy Alters Monocyte Differentiation to Favor Generation of Cancer-Supporting M2 Macrophages in the Tumor Microenvironment

RAS/PI3K Crosstalk and Cetuximab Resistance in Head and Neck Squamous Cell Carcinoma

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