Evaluation of cell-line-derived xenograft tumours as controls for immunohistochemical testing for ER and PR

2015 ◽  
Vol 68 (9) ◽  
pp. 746-751 ◽  
Author(s):  
Tahrim Hasan ◽  
Beverley Carter ◽  
Nash Denic ◽  
Luis Gai ◽  
Jennifer Power ◽  
...  
Keyword(s):  
Quality Control ◽  
Cell Line ◽  
Cell Lines ◽  
Control Material ◽  
Biomarker Testing ◽  
Steady State Levels ◽  
Archived Specimens ◽  
Different Levels ◽  
Immunohistochemical Testing ◽  
Fat Pads

Quality control (QC) for immunohistochemistry (IHC) analysis routinely incorporates archived specimens for on-slide control material. We have assessed the utility of cell-line-derived xenograft (CDX) tumours for QC in breast estrogen receptor (ER) and progesterone receptor (PR) biomarker testing. Immunoblot and IHC analyses were used to select cell lines with different steady-state levels of ER and PR expression. CDX tumours all demonstrated consistent and comparable expression of ER and PR with corresponding cell lines from which they were derived. Three pathologists experienced in breast biomarker reporting scored tumours from different locations on mammary fat pads to determine reproducibility. Tumours from different locations were consistently scored as identical, and the CDX tumours representing different levels of biomarker expression were similar to patient-derived controls. Pathologists could not consistently distinguish CDX tumours from patient-derived controls, suggesting that within the appropriate quality management setting, CDX tumours may serve as control material for reporting purposes.

scholarly journals Keeping it clean: the cell culture quality control experience at the National Center for Advancing Translational Sciences

2019 ◽  
Author(s):  
Jacob S. Roth ◽  
Tobie D. Lee ◽  
Dorian M. Cheff ◽  
Maya L. Gosztyla ◽  
Rosita Asawa ◽  
...  
Keyword(s):  
Quality Control ◽  
Cell Line ◽  
Cell Lines ◽  
High Throughput Screening ◽  
Science Research ◽  
Rapid Identification ◽  
Contamination Rate ◽  
Mycoplasma Contamination ◽  
Systematic Testing ◽  
Short Tandem

AbstractQuality control monitoring of cell lines utilized in biomedical research is of critical importance, and is critical for reproducibility of data. Two key pitfalls in tissue culture are 1) cell line authenticity, and 2) mycoplasma contamination. As a collaborative research institute, the National Center for Advancing Translational Sciences receives cell lines from a range of commercial and academic sources, that are adapted for high-throughput screening. Here, we describe the implementation of a routine NCATS-wide mycoplasma testing and short-tandem repeat (STR) testing for cell lines. Initial testing identified a >10% mycoplasma contamination rate, but the implementation of clearly defined protocols that included immediate destruction of contaminated cell lines wherever possible has led to a much-reduced mycoplasma contamination rate, and data for >2,000 cell line samples tested over 3 years, and case studies are provided. STR testing of 170 cell lines with established STR profiles revealed only 5 mis-identified cell lines received from external labs. The data collected over the three years since implementation of this systematic testing demonstrates the importance of continual vigilance for rapid identification of ‘problem’ cell lines, for ensuring reproducible data in translational science research.

scholarly journals Comprehensive Profiling of Surface Gangliosides Extracted from Various Cell Lines by LC-MS/MS

Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1323
Author(s):  
Jua Lee ◽  
Heeyoun Hwang ◽  
Sumin Kim ◽  
Jaeyun Hwang ◽  
Jaekyung Yoon ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cell Types ◽  
Cellular Membrane ◽  
Abundant Species ◽  
Invasion And Metastasis ◽  
Pancreatic Cell ◽  
Analytical Tools ◽  
Different Levels

Gangliosides act as a surface marker at the outer cellular membrane and play key roles in cancer cell invasion and metastasis. Despite the biological importance of gangliosides, they have been still poorly characterized due to the lack of effective analytical tools. Herein, we performed molecular profiling and structural elucidation of intact gangliosides in various cell lines including CFPAC1, A549, NCI-H358, MCF7, and Caski. We identified and quantified a total of 76 gangliosides on cell membrane using C18 LC-MS/MS. Gangliosides found in each cell line exhibited high complexity and diversity both qualitatively and quantitatively. The most abundant species was GM3(d34:1) in CFPAC1, NCI-H358, and MCF7, while GM2(d34:1) and GM1(d34:1) were major components in A549 and Caski, respectively. Notably, glycan moieties showed more diversity between cancer cell lines than ceramide moieties. In addition, noncancerous pancreatic cell line (hTERT/HPNE) could be distinguished by gangliosides containing different levels of sialic acid compared with cancerous pancreatic cell line (CFPAC1). These results clearly demonstrated the feasibility of our analytical platform to comprehensive profile of cell surface gangliosides for identifying cell types and subgrouping cancer cell types.

scholarly journals Rapid and Quantitative Assessment of Cell Quality, Identity, and Functionality for Cell-Based Assays Using Real-Time Cellular Analysis

2011 ◽  
Vol 16 (3) ◽  
pp. 313-322 ◽  
Author(s):  
Jeffrey T. Irelan ◽  
Meng-Jou Wu ◽  
Jonathan Morgan ◽  
Ning Ke ◽  
Biao Xi ◽  
...  
Keyword(s):  
Quality Control ◽  
Cell Line ◽  
Real Time ◽  
Cell Lines ◽  
Quantitative Measure ◽  
Genetic Alterations ◽  
Cellular Analysis ◽  
Identical Cell ◽  
Reproducible Method ◽  
Impedance Profile

Strict quality control of cells is required for the standardization and interpretation of results in all areas of cell-based research, especially in drug discovery. Real-time cellular analysis using electrical impedance as a readout offers a rapid and highly reproducible method for quality control as it provides a quantitative measure of overall cell morphology and growth. In a case study, the authors demonstrate that samples of a single cell line obtained from several different labs show clear differences in their impedance profiles when compared with the corresponding standard cell line. A number of kinetic parameters were derived from the impedance profiles and used to quantify the differences among these cell lines. Our findings indicate that this methodology can detect cell line differences including mix-ups or contaminations, genetic alterations, and potential epigenetic changes occurring during passaging, all of which can occur in the time scale of a screening campaign. Finally, we provide evidence that these impedance profile differences can be predictive of different outcomes in cell-based functional assays for the effects of small molecules on otherwise seemingly identical cell lines.

scholarly journals Keeping It Clean: The Cell Culture Quality Control Experience at the National Center for Advancing Translational Sciences

2020 ◽  
Vol 25 (5) ◽  
pp. 491-497 ◽  
Author(s):  
Jacob S. Roth ◽  
Tobie D. Lee ◽  
Dorian M. Cheff ◽  
Maya L. Gosztyla ◽  
Rosita R. Asawa ◽  
...  
Keyword(s):  
Quality Control ◽  
Cell Line ◽  
Cell Lines ◽  
High Throughput Screening ◽  
Science Research ◽  
Rapid Identification ◽  
Contamination Rate ◽  
Mycoplasma Contamination ◽  
Systematic Testing ◽  
Short Tandem

Quality control monitoring of cell lines utilized in biomedical research is of utmost importance and is critical for the reproducibility of data. Two key pitfalls in tissue culture are 1) cell line authenticity and 2) Mycoplasma contamination. As a collaborative research institute, the National Center for Advancing Translational Sciences (NCATS) receives cell lines from a range of commercial and academic sources, which are adapted for high-throughput screening. Here, we describe the implementation of routine NCATS-wide Mycoplasma testing and short tandem repeat (STR) testing for cell lines. Initial testing identified a >10% Mycoplasma contamination rate. While the implementation of systematic testing has not fully suppressed Mycoplasma contamination rates, clearly defined protocols that include the immediate destruction of contaminated cell lines wherever possible has enabled rapid intervention and removal of compromised cell lines. Data for >2000 cell line samples tested over 3 years, and case studies are provided. STR testing of 186 cell lines with established STR profiles revealed only five misidentified cell lines, all of which were received from external labs. The data collected over the 3 years since implementation of this systematic testing demonstrate the importance of continual vigilance for rapid identification of “problem” cell lines, for ensuring reproducible data in translational science research.

scholarly journals A novel cell line generated using the CRISPR/Cas9 technology as universal quality control material for KRAS G12V mutation testing

2018 ◽  
Vol 32 (5) ◽  
pp. e22391 ◽  
Author(s):  
Shiyu Jia ◽  
Rui Zhang ◽  
Guigao Lin ◽  
Rongxue Peng ◽  
Peng Gao ◽  
...  
Keyword(s):  
Quality Control ◽  
Cell Line ◽  
Mutation Testing ◽  
Quality Control Material ◽  
Control Material

scholarly journals Efficient and crucial quality control of HAP1 cell ploidy status

Biology Open ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. bio057174 ◽  
Author(s):  
Tobias B. Beigl ◽  
Ine Kjosås ◽  
Emilie Seljeseth ◽  
Nina Glomnes ◽  
Henriette Aksnes
Keyword(s):  
Flow Cytometry ◽  
Quality Control ◽  
Cell Line ◽  
Cell Lines ◽  
Human Cell ◽  
Cell Sorting ◽  
Gene Editing ◽  
Human Cell Line ◽  
Popular Subject ◽  
Ploidy Status

ABSTRACTThe near-haploid human cell line HAP1 recently became a popular subject for CRISPR/Cas9 editing, since only one allele requires modification. Through the gene-editing service at Horizon Discovery, there are at present more than 7500 edited cell lines available and the number continuously increases. The haploid nature of HAP1 is unstable as cultures become diploid with time. Here, we demonstrated some fundamental differences between haploid and diploid HAP1 cells, hence underlining the need for taking control over ploidy status in HAP1 cultures prior to phenotyping. Consequently, we optimized a procedure to determine the ploidy of HAP1 by flow cytometry in order to obtain diploid cultures and avoid ploidy status as an interfering variable in experiments. Furthermore, in order to facilitate this quality control, we validated a size-based cell sorting procedure to obtain the diploid culture more rapidly. Hence, we provide here two streamlined protocols for quality controlling the ploidy of HAP1 cells and document their validity and necessity.This article has an associated First Person interview with the co-first authors of the paper.

scholarly journals CRITERION-ORIENTED TEST BASED ON THE USE OF COMPUTERS AS A MEANS OF EVALUATING THE QUALITY OF STUDENTS’ KNOWLEDGE

2021 ◽  
pp. 81-97
Author(s):  
Dmitriy Prochukhan ◽  
Iryna Kostyria
Keyword(s):  
Higher Education ◽  
Quality Control ◽  
Computer Technology ◽  
Effective Means ◽  
Quality Characteristics ◽  
Test Results ◽  
Control Material ◽  
Professional Disciplines ◽  
Different Levels

The article considers new approaches to assessing student achievement in higher education. Modern priorities in education are considered, one of which is quality control of knowledge of future specialists. In general, mastering professional disciplines during training involves studying issues of application: designing tests of different levels of application, control material for use, interpretation of test results, interpretation of its results and drawing conclusions about its compliance with the purpose, objectives , understanding of the main quality characteristics of control material and the possibility of using computer technology in creating, conducting and processing test results. The article experimentally proves that assessing the quality of knowledge of futur e professionals using criterion-based testing based on the use of computers is one of the most effective means of pedagogical control.

The nuclear 3,5,3'-triiodothyronine receptor in human leukaemic cell lines

1984 ◽  
Vol 105 (3) ◽  
pp. 429-432 ◽  
Author(s):  
Juan Bernal ◽  
Leif C. Andersson
Keyword(s):  
Growth Hormone ◽  
Cell Line ◽  
Cell Lines ◽  
Rat Liver ◽  
Association Constant ◽  
Erythroid Differentiation ◽  
Leukaemic Cell ◽  
Cells In Culture ◽  
Gh Cells ◽  
High Concentrations

Abstract. The 3,5,3'-triiodothyronine (T3) receptor has been studied in a series of continuously growing human leukaemic cell lines. High concentrations of receptor were found in the erythroblastoid cell line K-562. T3 was bound to the nuclei of these cells with an association constant of 3.4 × 109 m−1, and capacity 104 fmol/100 μg DNA, or 8700 molecules/nucleus. This capacity is comparable to that of rat liver or growth hormone producing cells (GH cells) in culture, and suggests that the K-562 cell line could be a useful model for the study of T3 action on erythroid differentiation.

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