A solution to the generalized birthday problem with application to allozyme screening for cell culture contamination

1979 ◽  
Vol 16 (2) ◽  
pp. 242-251 ◽  
Author(s):  
M. H. Gail ◽  
G. H. Weiss ◽  
N. Mantel ◽  
S. J. O'Brien
Keyword(s):  
Cell Culture ◽  
Approximate Solution ◽  
Single Cell ◽  
Cell Line ◽  
General Problem ◽  
Birthday Problem ◽  
Classic Problem ◽  
Recursive Solution ◽  
Cell Culture Contamination

This paper gives the probability that no common allozyme signature is found among n randomly selected cell culture lines when the s mutually exclusive and exhaustive signatures have arbitrary known probabilities. This result is useful in detecting cell culture contamination by a single cell line. This problem is a generalization of the classic problem of finding the probability of no common birthday among n individuals when it is assumed that each individual has a chance 1/s = 1/365 of a birthday on a given day. Both an exact recursive solution and an approximate solution are given for the general problem.

A solution to the generalized birthday problem with application to allozyme screening for cell culture contamination

1979 ◽  
Vol 16 (02) ◽  
pp. 242-251 ◽  
Author(s):  
M. H. Gail ◽  
G. H. Weiss ◽  
N. Mantel ◽  
S. J. O'Brien
Keyword(s):  
Cell Culture ◽  
Approximate Solution ◽  
Single Cell ◽  
Cell Line ◽  
General Problem ◽  
Birthday Problem ◽  
Classic Problem ◽  
Recursive Solution ◽  
Cell Culture Contamination

This paper gives the probability that no common allozyme signature is found among n randomly selected cell culture lines when the s mutually exclusive and exhaustive signatures have arbitrary known probabilities. This result is useful in detecting cell culture contamination by a single cell line. This problem is a generalization of the classic problem of finding the probability of no common birthday among n individuals when it is assumed that each individual has a chance 1/s = 1/365 of a birthday on a given day. Both an exact recursive solution and an approximate solution are given for the general problem.

scholarly journals Cell-Line Annotation on Europe PubMed Central

2014 ◽  
Author(s):  
Jee-Hyub Kim
Keyword(s):  
Cell Culture ◽  
Single Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Biomedical Literature ◽  
Pubmed Central ◽  
Preliminary Results ◽  
Data Citation ◽  
A Cell

A cell line is a cell culture developed from a single cell and therefore consisting of cells with a uniform genetic make-up. A cell line has an important role as a research resource such as organisms, antibodies, constructs, knockdown reagents, etc. Unique identification of cell lines in the biomedical literature is important for the reproducibility of science. As data citation, resource citation is also important for resource re-use. In this paper, we mention the challenges of identifying cell lines and describe a system for cell line annotation with preliminary results.

Henrietta Lacks, HeLa Cells, and Cell Culture Contamination

2009 ◽  
Vol 133 (9) ◽  
pp. 1463-1467 ◽  
Author(s):  
Brendan P. Lucey ◽  
Walter A. Nelson-Rees ◽  
Grover M. Hutchins
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Cell Line ◽  
Hela Cells ◽  
Human Cell Line ◽  
Hela Cell Line ◽  
Cell Line Hela ◽  
The World ◽  
Cell Culture Contamination ◽  
Line Hela

Abstract Henrietta Lacks died in 1951 of an aggressive adenocarcinoma of the cervix. A tissue biopsy obtained for diagnostic evaluation yielded additional tissue for Dr George O. Gey's tissue culture laboratory at Johns Hopkins (Baltimore, Maryland). The cancer cells, now called HeLa cells, grew rapidly in cell culture and became the first human cell line. HeLa cells were used by researchers around the world. However, 20 years after Henrietta Lacks' death, mounting evidence suggested that HeLa cells contaminated and overgrew other cell lines. Cultures, supposedly of tissues such as breast cancer or mouse, proved to be HeLa cells. We describe the history behind the development of HeLa cells, including the first published description of Ms Lacks' autopsy, and the cell culture contamination that resulted. The debate over cell culture contamination began in the 1970s and was not harmonious. Ultimately, the problem was not resolved and it continues today. Finally, we discuss the philosophical implications of the immortal HeLa cell line.

In vitro Three Dimensional Scaffold-free Construct of Human Adipose-derived Stem Cells in Coculture with Endothelial Cells and Fibroblasts

2017 ◽  
Vol 68 (6) ◽  
pp. 1341-1344
Author(s):  
Grigore Berea ◽  
Gheorghe Gh. Balan ◽  
Vasile Sandru ◽  
Paul Dan Sirbu
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Endothelial Cells ◽  
Single Cell ◽  
Cell Line ◽  
Bone Repair ◽  
Umbilical Vein ◽  
Human Fibroblasts ◽  
Three Dimensional ◽  
Adipose Derived Stem Cells

Complex interactions between stem cells, vascular cells and fibroblasts represent the substrate of building microenvironment-embedded 3D structures that can be grafted or added to bone substitute scaffolds in tissue engineering or clinical bone repair. Human Adipose-derived Stem Cells (hASCs), human umbilical vein endothelial cells (HUVECs) and normal dermal human fibroblasts (NDHF) can be mixed together in three dimensional scaffold free constructs and their behaviour will emphasize their potential use as seeding points in bone tissue engineering. Various combinations of the aforementioned cell lines were compared to single cell line culture in terms of size, viability and cell proliferation. At 5 weeks, viability dropped for single cell line spheroids while addition of NDHF to hASC maintained the viability at the same level at 5 weeks Fibroblasts addition to the 3D construct of stem cells and endothelial cells improves viability and reduces proliferation as a marker of cell differentiation toward osteogenic line.

An artificial intelligent platform for live cell identification and the detection of cross-contamination (Preprint)

2019 ◽  
Author(s):  
Ruixin Wang ◽  
Dongni Wang ◽  
Dekai Kang ◽  
Xusen Guo ◽  
Chong Guo ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Cell Morphology ◽  
Single Cell Level ◽  
Cross Contamination ◽  
Cell Level ◽  
Cell Identification ◽  
Pure Cell ◽  
Microscopy Images

BACKGROUND In vitro human cell line models have been widely used for biomedical research to predict clinical response, identify novel mechanisms and drug response. However, one-fifth to one-third of cell lines have been cross-contaminated, which can seriously result in invalidated experimental results, unusable therapeutic products and waste of research funding. Cell line misidentification and cross-contamination may occur at any time, but authenticating cell lines is infrequent performed because the recommended genetic approaches are usually require extensive expertise and may take a few days. Conversely, the observation of live-cell morphology is a direct and real-time technique. OBJECTIVE The purpose of this study was to construct a novel computer vision technology based on deep convolutional neural networks (CNN) for “cell face” recognition. This was aimed to improve cell identification efficiency and reduce the occurrence of cell-line cross contamination. METHODS Unstained optical microscopy images of cell lines were obtained for model training (about 334 thousand patch images), and testing (about 153 thousand patch images). The AI system first trained to recognize the pure cell morphology. In order to find the most appropriate CNN model，we explored the key image features in cell morphology classification tasks using the classical CNN model-Alexnet. After that, a preferred fine-grained recognition model BCNN was used for the cell type identification (seven classifications). Next, we simulated the situation of cell cross-contamination and mixed the cells in pairs at different ratios. The detection of the cross-contamination was divided into two levels, whether the cells are mixed and what the contaminating cell is. The specificity, sensitivity, and accuracy of the model were tested separately by external validation. Finally, the segmentation model DialedNet was used to present the classification results at the single cell level. RESULTS The cell texture and density were the influencing factors that can be better recognized by the bilinear convolutional neural network (BCNN) comparing to AlexNet. The BCNN achieved 99.5% accuracy in identifying seven pure cell lines and 86.3% accuracy for detecting cross-contamination (mixing two of the seven cell lines). DilatedNet was applied to the semantic segment for analyzing in single-cell level and achieved an accuracy of 98.2%. CONCLUSIONS This study successfully demonstrated that cell lines can be morphologically identified using deep learning models. Only light-microscopy images and no reagents are required, enabling most labs to routinely perform cell identification tests.

scholarly journals Single-cell RNA sequencing of human fetal epicardium reveals novel markers and regulators of EMT

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
T.J Streef ◽  
T Van Herwaarden ◽  
A.M Smits ◽  
M.J Goumans
Keyword(s):  
Cell Culture ◽  
Single Cell ◽  
Rna Sequencing ◽  
Public Institution ◽  
Funding Source ◽  
Injury Response ◽  
Post Injury ◽  
Paracrine Factors ◽  
Adult Heart ◽  
Single Cell Rna Sequencing

Abstract Background The heart is covered by the epicardium, consisting of epithelial cells and a mesenchymal layer. The epicardium has been shown to be essential during cardiac development by contributing cells through epithelial-to-mesenchymal transition (EMT) and the secretion of paracrine factors. In the adult, the epicardium conveys a cardioprotective response after myocardial infarction, albeit suboptimal compared to the epicardial contribution to heart development. Although the developing epicardium has been characterised in mice and zebrafish, knowledge on the human fetal epicardium derives mostly from cell culture models. Therefore, direct analysis of the human fetal epicardium is vital as it provides new insights into the cellular and biochemical interactions within the developing heart, which can potentially contribute to enhancing the post-injury response. Aim To study the human fetal epicardium using single-cell RNA sequencing (scRNA seq) in order to determine its cellular compositionThe data are further explored to e.g.identify regulators of epicardial EMT. Methods Epicardial layers were isolated from four fetal human hearts (14–15 weeks gestation, obtained under informed consent and according to local ethical approval). Tissue was digested, and single live cells were sorted into 384-wells plates and sequenced. Data analysis was performed using R-packages RaceID3 and StemID2. Findings were validated using qPCR and immunohistochemistry. Results Analysis of 2024 cells reveals a clear clustering of the epicardial epithelium and the mesenchymal population. Importantly, we found that “classical” markers, such as Wilms' Tumor 1 and T-box transcription factor 18, are not specific enough to reliably identify the epicardium, but our analysis has provided markers that do allow for robust identification of the epicardium. Additionally, we were able to identify epicardial subpopulations based on their expression profile, and we are currently investigating these using immunohistochemistry in human fetal and adult heart tissue sections. To establish the regulation of epicardial activation we are focussing on the process of EMT within our dataset using RaceID2. From our analysis, several regulators of epicardial EMT are proposed that will be followed up on in vitro. Conclusions We identify various novel markers of the fetal epithelial epicardium, as well as characterizing markers of the mesenchymal layer. We also identified novel factors involved in epicardial EMT, and these are currently being validated in our cell-culture model. These data can provide new insights into the post-injury response in the adult heart. Funding Acknowledgement Type of funding source: Public Institution(s). Main funding source(s): Dutch Heart Foundation

scholarly journals Tribological, biocompatibility, and antibiofilm properties of tungsten–germanium coating using magnetron sputtering

2021 ◽  
Vol 32 (1) ◽  
Author(s):  
Mustafa Şükrü Kurt ◽  
Mehmet Enes Arslan ◽  
Ayşenur Yazici ◽  
İlkan Mudu ◽  
Elif Arslan
Keyword(s):  
Cell Culture ◽  
Magnetron Sputtering ◽  
Cell Line ◽  
Borosilicate Glass ◽  
Chromosomal Abnormalities ◽  
Dermal Fibroblast ◽  
Fibroblast Cell ◽  
Antibiofilm Activity ◽  
Borosilicate Glasses

AbstractIn this study, borosilicate glass and 316 L stainless steel were coated with germanium (Ge) and tungsten (W) metals using the Magnetron Sputtering System. Surface structural, mechanical, and tribological properties of uncoated and coated samples were examined using SEM, X-ray diffraction (XRD), energy-dispersive spectroscopy, and tribometer. The XRD results showed that WGe2 chemical compound observed in (110) crystalline phase and exhibited a dense structure. According to the tribological analyses, the adhesion strength of the coated deposition on 316 L was obtained 32.8 N, and the mean coefficient of friction was around 0.3. Biocompatibility studies of coated metallic biomaterials were analyzed on fibroblast cell culture (Primary Dermal Fibroblast; Normal, Human, Adult (HDFa)) in vitro. Hoescht 33258 fluorescent staining was performed to investigate the cellular density and chromosomal abnormalities of the HDFa cell line on the borosilicate glasses coated with germanium–tungsten (W–Ge). Cell viabilities of HDFa cell line on each surface (W–Ge coated borosilicate glass, uncoated borosilicate glass, and cell culture plate surface) were analyzed by using (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cytotoxicity assay. The antibiofilm activity of W–Ge coated borosilicate glass showed a significant reduction effect on Staphylococcus aureus (ATCC 25923) and Pseudomonas aeruginosa (ATCC 27853) adherence compared to control groups. In the light of findings, tungsten and germanium, which are some of the most common industrial materials, were investigated as biocompatible and antimicrobial surface coatings and recommended as bio-implant materials for the first time.

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