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G3 (Bethesda, Md.) Aug 2013HeLa is the most widely used model cell line for studying human cellular and molecular biology. To date, no genomic reference for this cell line has been released, and...
HeLa is the most widely used model cell line for studying human cellular and molecular biology. To date, no genomic reference for this cell line has been released, and experiments have relied on the human reference genome. Effective design and interpretation of molecular genetic studies performed using HeLa cells require accurate genomic information. Here we present a detailed genomic and transcriptomic characterization of a HeLa cell line. We performed DNA and RNA sequencing of a HeLa Kyoto cell line and analyzed its mutational portfolio and gene expression profile. Segmentation of the genome according to copy number revealed a remarkably high level of aneuploidy and numerous large structural variants at unprecedented resolution. Some of the extensive genomic rearrangements are indicative of catastrophic chromosome shattering, known as chromothripsis. Our analysis of the HeLa gene expression profile revealed that several pathways, including cell cycle and DNA repair, exhibit significantly different expression patterns from those in normal human tissues. Our results provide the first detailed account of genomic variants in the HeLa genome, yielding insight into their impact on gene expression and cellular function as well as their origins. This study underscores the importance of accounting for the strikingly aberrant characteristics of HeLa cells when designing and interpreting experiments, and has implications for the use of HeLa as a model of human biology.
Topics: Alleles; DNA Copy Number Variations; Databases, Genetic; Gene Frequency; Genome, Human; Genomics; HeLa Cells; Humans; Models, Biological; Mutation; RNA Interference; Sequence Analysis, DNA; Sequence Analysis, RNA; Transcriptome
PubMed: 23550136
DOI: 10.1534/g3.113.005777 -
G3 (Bethesda, Md.) Jun 2013
Topics: Genetic Privacy; Genome, Human; HeLa Cells; Humans; Informed Consent; Sequence Analysis, DNA
PubMed: 23576522
DOI: 10.1534/g3.113.006304 -
Journal of Cellular and Molecular... Jan 2019HeLa cells are a commonly used cell line in many biological research areas. They are not picky for culture medium and proliferate rapidly. HeLa cells are a notorious...
HeLa cells are a commonly used cell line in many biological research areas. They are not picky for culture medium and proliferate rapidly. HeLa cells are a notorious source of cell cross-contamination and have been found to be able to contaminate a wide range of cell lines in cell culture. In this study, we reported a simple and efficient method for detecting the presence of HeLa cell contamination in cell culture. HPV-18 was used as a biomarker. The cell culture supernatant was used directly as the template for nested PCR without extracting nucleic acid. By PCR amplification of the cell culture supernatant with the designed primers, we were able to detect the presence of HeLa cells in the culture. The sensitivity of this method can reach 1%, which is 10-fold higher than Short tandem repeat sequence (STR) profiling. This simple, rapid, and "noninvasive" quality checking method should find applications in routine cell culture practice.
Topics: Cell Culture Techniques; Cell Line; HeLa Cells; Human papillomavirus 18; Humans; Microsatellite Repeats; Polymerase Chain Reaction; Reproducibility of Results
PubMed: 30353657
DOI: 10.1111/jcmm.13923 -
Scientific Reports Oct 2019We have characterized site-specific N-glycosylation of the HeLa cell line glycoproteins, using a complex workflow based on high and low energy tandem mass spectrometry...
We have characterized site-specific N-glycosylation of the HeLa cell line glycoproteins, using a complex workflow based on high and low energy tandem mass spectrometry of glycopeptides. The objective was to obtain highly reliable data on common glycoforms, so rigorous data evaluation was performed. The analysis revealed the presence of a high amount of bovine serum contaminants originating from the cell culture media - nearly 50% of all glycans were of bovine origin. Unaccounted, the presence of bovine serum components causes major bias in the human cellular glycosylation pattern; as is shown when literature results using released glycan analysis are compared. We have reliably identified 43 (human) glycoproteins, 69 N-glycosylation sites, and 178 glycoforms. HeLa glycoproteins were found to be highly (68.7%) fucosylated. A medium degree of sialylation was observed, on average 46.8% of possible sialylation sites were occupied. High-mannose sugars were expressed in large amounts, as expected in the case of a cancer cell line. Glycosylation in HeLa cells is highly variable. It is markedly different not only on various proteins but also at the different glycosylation sites of the same protein. Our method enabled the detailed characterization of site-specific N-glycosylation of several glycoproteins expressed in HeLa cell line.
Topics: Culture Media; Glycoproteins; Glycosylation; HeLa Cells; Humans; Metabolomics; Polysaccharides; Tandem Mass Spectrometry
PubMed: 31616032
DOI: 10.1038/s41598-019-51428-x -
Archives of Razi Institute Oct 2022The selection of suitable cell cultures for use in the biological industry as well as research and diagnostic studies is critical. One of the factors affecting cell...
The selection of suitable cell cultures for use in the biological industry as well as research and diagnostic studies is critical. One of the factors affecting cell culture that can affect the results of studies is the contamination with viral agents. Therefore, efforts to preserve the health of cell cultures from contamination sound logical, and the use of virus-free cells is vital in research and diagnostic studies as well as in the manufacturing industries. For this purpose, it is crucial to use fast and correct diagnostic methods to detect the presence of critical viral contaminants in cells. Moreover, the use of susceptible diagnostic methods is also doubly important, especially in the case of contaminants that may remain hidden. Therefore, in this study, the BHK-21C5 cell line, one of the most widely used cells in the production and quality control of biological products and virological studies, was examined in terms of contamination with the most important viruses such as BVD and BLV. Detection of possible contaminants by using two-step RT-PCR to detect the 5' UTR portion of the BVD virus. Moreover, Nested PCR was carried out to detect the BLV virus using the gp51 gene region. Also, an Flk cell line and Hela cell line that were consistently contaminated with the BLV virus were used as positive controls in Nested PCR. Due to the absence of bands in the BHK-21C5 cell column and the bandwidth observed in the positive control column (BVDV) in the range of 283 bp, non-contaminating of the cell clone with the BVD virus was proved. Also, no band was observed in well related to BHK-21C5 cell, and no cell clone contamination with BLV was confirmed, and in wells related to the positive controls (Flk and Hela cells), the bands have seen in the range 444bp. So, the results showed that no obvious or covert viral contamination effect was observed in the cell clone studied. Hence, the use of this cell line seems unobstructed in the quality control and production of biological products and research and diagnostic studies.
Topics: Animals; Humans; HeLa Cells; Polymerase Chain Reaction; Viruses; Cricetinae
PubMed: 37123112
DOI: 10.22092/ARI.2021.356543.1867 -
Medecine Sciences : M/S Dec 2021Many developments in biology and biotechnology have relied on the HeLa cell line, originally derived in 1951 from a Black cancer patient without her knowledge. This...
Many developments in biology and biotechnology have relied on the HeLa cell line, originally derived in 1951 from a Black cancer patient without her knowledge. This origin became generally known at the turn of the century, and the patient's descendants have sought and obtained some recognition and some control but little compensation. They have now retained two famous attorneys to sue a biotech firm for very extensive damages, with more legal action planned against other companies. This may have important repercussions for the biotech industry, and raises complex issues regarding ownership of biological material and compensation to patients from whom these materials have been obtained.
Topics: HeLa Cells; Humans
PubMed: 34928229
DOI: 10.1051/medsci/2021181 -
Le Infezioni in Medicina Jun 2018Toxoplasma gondii is a protozoan parasite which can be grown in vivo and in vitro. Various cell lines are used for T. gondii culture in vitro. In this study, four cell...
Toxoplasma gondii is a protozoan parasite which can be grown in vivo and in vitro. Various cell lines are used for T. gondii culture in vitro. In this study, four cell lines of HeLa, Vero, RBK and A549 were compared with each other for T. gondii tachyzoites culture. The four cell lines were cultured and infected with 5,000,000 tachyzoites, respectively. The number of tachyzoites and viable host cells and pH of the media were assessed in each culture. The highest tachyzoite yield was seen in HeLa cell culture. The lowest number of viable host cells and the lowest pH were seen in HeLa cell line culture. The lowest tachyzoite yield, the highest viable cell and the highest pH were observed in Vero cell line culture. HeLa and Vero cell lines are thus appropriate for rapid and long-term propagations of T. gondii tachyzoites, respectively.
Topics: A549 Cells; Animals; Chlorocebus aethiops; HeLa Cells; Humans; Parasitology; Toxoplasma; Vero Cells
PubMed: 29932087
DOI: No ID Found -
BMC Complementary Medicine and Therapies Aug 2023Cervical cancer remains a significant global health issue, highlighting the need for effective therapeutic strategies. Given that Sphaerocoryne affinis (SA) has shown...
BACKGROUND
Cervical cancer remains a significant global health issue, highlighting the need for effective therapeutic strategies. Given that Sphaerocoryne affinis (SA) has shown potential anti-cancer activity in several cancer types, herein, we investigate the effects of SA fruit (SAF) on human cervical cancer HeLa cells and their underlying mechanisms of action.
METHODS
SAF extract cytotoxicity was assessed in various cancer cell lines. The effects of the hexane fraction (SAF-Hex) on HeLa cell viability, cell cycle protein expression, apoptosis, and DNA damage were evaluated using cytotoxicity assays, Western blotting, quantitative PCR, 4',6-diamidino-2-phenylindole (DAPI) staining, and a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay.
RESULTS
SAF-Hex selectively inhibited HeLa cell viability with an IC50 of 4.20 ± 0.36 µg/mL and a selectivity index of 5.11 ± 0.58. The time-dependent cytotoxicity assay showed decreased cell survival after 48 h of treatment, accompanied by morphological changes and apoptotic bodies in HeLa cells. SAF-Hex also suppressed HeLa cell cycle proteins (Cyclin E, CDK2, and CDK1), reduced PCNA transcription, and diminished AKT and mTOR activation, thus inhibiting cell proliferation. The increased γH2AX expression, DNA fragmentation, and caspases-3 and -9 activation indicated SAF-Hex-induced DNA damage and apoptosis. However, the BAX/BCL-2 ratio remained unchanged, and BAX and BCL2 expression was attenuated.
CONCLUSION
SAF-Hex effectively inhibits HeLa cell proliferation and induces DNA damage in that cervical cancer cell line activating apoptosis through the intrinsic pathway. Interestingly, the BAX/BCL-2 ratio remained unchanged while BAX and BCL2 transcription was attenuated. Hence, further research is required to explore this unexpected finding and facilitate the development of novel therapies targeting cervical cancer HeLa cells.
Topics: Humans; Female; Uterine Cervical Neoplasms; HeLa Cells; Fruit; bcl-2-Associated X Protein; Apoptosis
PubMed: 37598145
DOI: 10.1186/s12906-023-04127-0 -
BMC Research Notes Jun 2018Cell density in tumor cell three dimensional (3D) cultures affects secretome expression of components. A microenvironment characteristic shared by high-density 3D cell...
OBJECTIVE
Cell density in tumor cell three dimensional (3D) cultures affects secretome expression of components. A microenvironment characteristic shared by high-density 3D cell culture and in vivo tumor masses is poor oxygenation, with anoxia being a natural cell state in tumor centers. Until recently, the ability to study anoxia-adapted cell physiology was not possible. Using a newly-developed methodology, anoxic HeLa cell secretome expression was measured.
RESULTS
Anoxic HeLa cell cytokine levels after 3 days' (hypoxia inducible factor, HIF1 positive) and 10 days' growth (HIF1 negative; anaerobic respiration) were significantly (p < 0.01) higher than normoxic controls for: IL-8 (1.8- and 3.4-fold higher, respectively), GRO (1.3- and 1.1-fold higher, respectively), and IL-11 (1.4- and 1.1-fold higher, respectively). In contrast, G-CSF, IFNα2, and CXCL-10 levels decreased over time (day 3 vs. day 10). Thus, metabolically active HeLa cells respond to the lack of oxygen, in part, by regulating the levels of cytokines produced. Cytokines expressed at increased levels, in the absence of oxygen, correspond to a secretomic profile reported for paracrine signaling pathways associated with metastasis. Further studies defining physiologic changes that occur upon anoxic growth may lead to the discovery of novel chemotherapeutic drug targets.
Topics: Cell Hypoxia; Cytokines; Gene Expression; HeLa Cells; Humans; Neoplasms; Oxygen
PubMed: 29941048
DOI: 10.1186/s13104-018-3520-5 -
Microbiology (Reading, England) Dec 2002The interactions of group B streptococci (GBS) with HeLa cells (an epithelial cell line) and MRC-5 cells (a fibroblastic cell line) were explored. A host-cell invasion...
The interactions of group B streptococci (GBS) with HeLa cells (an epithelial cell line) and MRC-5 cells (a fibroblastic cell line) were explored. A host-cell invasion assay using GBS strains from all serotypes revealed that GBS invaded HeLa cells to a greater extent than MRC-5 cells. One strain, a serotype V (NCS13), was highly invasive against HeLa cells. All strains were poorly invasive against MRC-5 cells. Further characterization of the binding of NCS13 to HeLa and MRC-5 cell surfaces showed that the lack of recoverable c.f.u. from MRC-5 cells was due to a lack of binding of NCS13 to the MRC-5 cell surface in comparison to HeLa cells. Although fibronectin had been reported to bind to GBS, fibronectin assays showed 2.7-fold more fibronectin on the MRC-5 cell surface in comparison to HeLa cells, suggesting that other extracellular matrix proteins besides fibronectin may be involved in GBS binding. Scanning electron microscopy of NCS13 and HeLa cells over a 6 h time period showed increased numbers of NCS13 on the HeLa cell surface over time until cell death at 6 h. Direct contact of the HeLa cell surface by NCS13 was found to be necessary for cell death to occur. Further scanning electron microscopy studies found that, once GBS are bound to the HeLa cell surface, HeLa cell microvilli entwine the bacteria, which then enter the HeLa cell in a polar fashion. Cytoskeletal actin is involved, as this process is disrupted by cytochalasin D, and recruitment of actin is visible at the site of adherent chains of GBS. Also, the host-cell signalling enzyme, PI 3-kinase, is involved in the GBS internalization process, since the PI 3-kinase inhibitor, wortmannin, inhibited NCS13 invasion of HeLa cells in a dose-dependent manner.
Topics: Actins; Adult; Aged; Aged, 80 and over; Bacterial Adhesion; Cell Line; Cytochalasin D; Fibroblasts; Fibronectins; HeLa Cells; Humans; Infant; Infant, Newborn; Microscopy, Electron, Scanning; Middle Aged; Phosphatidylinositol 3-Kinases; Streptococcus agalactiae
PubMed: 12480896
DOI: 10.1099/00221287-148-12-3921