Use Of HeLa Cells Research Articles

Descendants of Henrietta Lacks reach settlement with Thermo Fisher over its use of HeLa cells

Descendants of Henrietta Lacks reach settlement with Thermo Fisher over its use of HeLa cells

A Student-Led Hearing on the Use of HeLa Cells in Research

A Student-Led Hearing on the Use of HeLa Cells in Research

One-pot synthesis of gadolinium-doped carbon quantum dots for high-performance multimodal bioimaging.

In this study, an efficient and mild approach is reported for the facile synthesis of a carbon quantum dot (CQD)-based dual-modal fluorescence (FL)/magnetic resonance (MR) imaging probe by doping Gd(iii) into CQDs through a one-pot pyrolysis process at low temperature. The as-prepared Gd-encapsulated CQDs (GCDs), which have an average diameter of ∼15 nm, are highly water-soluble. The GCDs are observed to have a high MR response with a longitudinal relaxation of 57.42 mM-1 s-1 and a strong fluorescence brightness with an absolute quantum yield of 40% while containing only 1.0% (w/w) of Gd3+ content. Stemming from the minimum Gd-doping and the inert carbon coating, GCDs exhibit excellent biocompatibility and blood compatibility. Dual-modality bioimaging applications of GCDs are successfully demonstrated by the use of HeLa cells and mice as models, revealing their great potential in fundamental biomedical research studies and even clinical fields such as fluorescence-guided surgery and dual-modal FL/MR imaging of blood vessels.

Abstract 3463: The use of HeLa cells as a cancer genetics education tool for middle and high schools

Abstract Hands-on, inquiry based learning, and one-on-one mentoring are the most effective methods of disseminating scientific information pertaining to cancer. The Science Youth Initiative (SYI) is a K-12 science education partnership between the Louisiana State University Health Sciences Center in New Orleans (LSUHSC-NO) and local schools. The workshops are coordinated with LSUHSC-NO faculty and school teachers to maximize the level of student learning and complement their classroom curriculum and academic level. Since 2009, the SYI has reached more than 2100 K-12 students to date, of which about 77% were middle and high school students. A diverse group of trainees from the LSUHSC-NO Schools of Medicine, Graduate Studies, Public Health, Allied Health Professions, and Nursing help educate the K-12 students and serve as role models. The benefit of having a variety of health care professionals with a range of expertise is that students receive cancer information from multiple perspectives. The program utilizes the book, The Immortal Life of Henrietta Lacks by Rebecca Skloot, which is part of the curriculum in many schools. During these workshops, middle and high school students are able to stain and examine HeLa cell chromosomes, thereby learning about chromatin instability commonly associated with cancer. Students also learn about DNA isolation, PCR, restriction digestion, and gel electrophoresis as they engage in workshop discussions on science history, bioethics, the human papilloma virus (HPV), and cervical cancer. Pre and post workshop surveys were developed to evaluate the participants’ learning and retention of the material covered, self-confidence in the academic material, awareness of science career options, and evaluation of the program. Results of the current academic year demonstrate: (1) a 51% increase in test scores based on the material covered (2) 59% of students were interested in participating in a medical or research related internship after the workshop (3) 54% of students were interested in working part-time in the medical sciences after the workshop. Participating schools included rural, urban, public/charter and private schools. These varied in socioeconomic and ethnic backgrounds: some which had almost 100% African American/Hispanic student body, while others had a diverse distribution of African American, Hispanic, Asian, and Caucasian students. The program's use of HeLa cells as a cancer genetics education tool has been successful in making health sciences and cancer research topics more interesting and accessible, motivating a variety of students to improve their grades, and promoting various health science related career opportunities. Citation Format: Ayesha Umrigar, Mary Moore, Fern Tsien. The use of HeLa cells as a cancer genetics education tool for middle and high schools. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3463.

Loss of FADS2 function severely impairs the use of HeLa cells as an in vitro model for host response studies involving fatty acid effects.

ScopeEstablished epithelial cell lines equipped with pattern recognition receptors such as the Toll-like receptor (TLR)-2 are common tools for immune response studies on invading pathogens, e.g. the obligate intracellular species of Chlamydia. Moreover, such models are widely used to elucidate fatty acid-mediated immune effects. In several transformed cell lines, however, unusual loss of metabolic functions was described. The cell lines A549 and HeLa are poorly characterized in this respect. Therefore, we comparatively assessed the metabolic capacity of A549 and HeLa prior to proposed application as in vitro model for fatty acid effects on chlamydial infection.Methodology/Principal FindingsWe incubated both cell lines either with substrates (C18∶2n−6 or C18∶3n−3) or products (C18∶3n−6, C18∶4n−3) of fatty acid desaturase-2 (FADS2), and analysed the fatty acid profiles after 24 h and 72 h by gas chromatography. Based on these data, we suspected that the complete discontinuation of normal biosynthesis of long-chain polyunsaturated fatty acids (LC-PUFA) in HeLa was due to loss of FADS2 function. Consequently, prostaglandin E2 (PGE2) formation was less inducible by TLR2 stimulation in HeLa, likely as a result of not only insufficient supply of precursors but also weak cyclooxygenase-2 (COX-2) response. In accordance, Chlamydia infection rates were consistently lower in HeLa than in A549. Sequence analysis revealed no alteration within the FADS2 gene in HeLa. The FADS2 expression level, however, was significantly lower and, in contrast to A549, not regulated by C18∶2n−6. A549 exhibited regular fatty acid metabolism and enzyme functionality.Conclusions/SignificanceOur data show that HeLa cells considerably differ from A549 at several stages of fatty acid metabolism. The poor metabolic potential of HeLa, mainly concerning FADS2 upstream of COX-2 function, calls into question whether these cells represent a good model to unveil fatty acid or downstream eicosanoid effects in the course of intracellular bacterial infection.

Distribution of the Vasotocin Subtype Four Receptor (VT4R) in the Anterior Pituitary Gland of the Chicken, Gallus gallus, and its Possible Role in the Avian Stress Response

The neurohormone arginine vasotocin (AVT) in non mammalian vertebrates is homologous to arginine vasopressin (AVP) in mammals. Its actions are mediated via G protein-coupled receptors that belong to the vasotocin/mesotocin family. Because of the known regulatory effects of nonapeptide hormones on anterior pituitary functions, receptor subtypes in that family have been proposed to be located in anterior pituitary cells. Recently, an avian vasotocin receptor subtype designated VT4R has been cloned, which shares 69% sequence homology with a human vasopressin receptor, the V1aR. In the present study, a polyclonal antibody to the VT4R was developed and validated to confirm its specificity to the VT4R. The antibody was used to test the hypothesis that the VT4R is present in the avian anterior pituitary and is specifically associated with certain cell types, where its expression is modulated by acute stress. Western blotting of membrane protein extracts from pituitary tissue, the use of HeLa cells transfected with the VT4R and peptide competition assays all confirmed the specificity of the antibody to the VT4R. Dual-labelling immunofluorescence microscopy was utilised to identify pituitary cell types that contained immunoreactive VT4R. The receptor was found to be widely distributed throughout the cephalic lobe but not in the caudal lobe of the anterior pituitary. Immunoreactive VT4R was associated with corticotrophs. Approximately 89% of immunolabelled corticotrophs were shown to contain the VT4R. The immunoreactive VT4R was not found in gonadotrophs, somatotrophs or lactotrophs. To determine a possible functional role of the VT4R and previously characterised VT2R, gene expression levels in the anterior pituitary were determined after acute immobilisation stress by quantitative reverse transcriptase-polymerase chain reaction. The results showed a significant increase in plasma corticosterone levels (three- to four-fold), a significant reduction of VT4R mRNA and an increase of VT2R mRNA (P < 0.05) in acutely immobilised chicks compared to controls. The data suggest a role of the VT4R in the avian stress response.

Haunted by Henrietta: The Archive, Immortality and the Biological Arts

This article draws on Rebecca Schneider's thoughts on performance within archival culture (2001) in order to examine the use of HeLa cells in recent artistic practice. HeLa is one of the most commonly used cell lines in scientific research, renowned for its ‘immortality’ in vitro. The HeLa cell line originated from cervical cancer cells that were taken from an African American woman named Henrietta Lacks in 1951. Lacks died eight months after the biopsy, but her cells – which were removed without her knowledge and consent – live on and multiply in scientific laboratories worldwide. As artists working in the biological arts attempt to reimagine the controversial narratives surrounding HeLa and Henrietta Lacks, this paper argues that employing biological materials such as HeLa cells in artistic practice often gives rise to a flesh-like ‘object’ that resists the archive that instead privileges documentary and object remains. In this respect, the use of HeLa cells in some artistic work is reminiscent of performance's challenge to traditional archival logic. Despite disappearing in death, documentation and even in the moment of the live encounter, HeLa and its attendant histories nevertheless remain in material and immaterial acts that these artworks and their documentary texts participate in. Such practices importantly prompt areconceptualisation of our relationship to the body in light of the potential effects of contemporary biotechnological advancements on archival logic. It is this reconceptualisation which I suggest may be bestapproached, therefore, in dialogue with performance theory on appearance and disappearance within archival culture.

Multisite Phosphorylation of Nuclear Interaction Partner of ALK (NIPA) at G2/M Involves Cyclin B1/Cdk1

Nuclear interaction partner of ALK (NIPA) is an F-box-containing protein that defines a nuclear skp1 cullin F-box (SCF)-type ubiquitin E3 ligase (SCFNIPA) implicated in the regulation of mitotic entry. The SCFNIPA complex targets nuclear cyclin B1 for ubiquitination in interphase, whereas phosphorylation of NIPA in late G2 phase and mitosis inactivates the complex to allow for accumulation of cyclin B1. Here, we identify the region of NIPA that mediates binding to its substrate cyclin B1. In addition to the recently described serine residue 354, we specify 2 new residues, Ser-359 and Ser-395, implicated in the phosphorylation process at G2/M within this region. Moreover, we found cyclin B1/Cdk1 to phosphorylate NIPA at Ser-395 in mitosis. Mutation of both Ser-359 and Ser-395 impaired effective inactivation of the SCFNIPA complex, resulting in reduced levels of mitotic cyclin B1. These data are compatible with a process of sequential NIPA phosphorylation where cyclin B1/Cdk1 amplifies phosphorylation of NIPA once an initial phosphorylation event has dissociated the SCFNIPA complex. Thus, cyclin B1/Cdk1 may contribute to the regulation of its own abundance in early mitosis.

In vivo induced antigenic determinants ofActinobacillus actinomycetemcomitans

Actinobacillus actinomycetemcomitans is a Gram-negative capnophilic rod and the etiological agent of localized aggressive periodontitis. The genome-wide survey of A. actinomycetemcomitans using in vivo induced antigen technology (IVIAT) has previously resulted in the discovery of antigenic determinants expressed specifically in diseased patients. The present study evaluated the potential of these antigens as putative disease markers, and investigating their contribution to the pathogenesis of the microorganism. Sera from patients had a significantly greater antibody titer than sera from healthy controls against six antigens, which supports the in vivo expression of these antigens, and suggests their usefulness as disease markers. A. actinomycetemcomitans invasion of epithelium-derived HeLa cells resulted in the induction of all three genes tested, as evidenced by real-time PCR. Isogenic mutants of these three genes were constructed and the adhesion and intracellular survival of the mutants was assayed in a competition assay with the wild-type strain. A significant defect in the intracellular survival of two of these mutant strains (orf1402 and orf859) was found. This defect could not be attributed to an adhesion defect. In contrast, a mutation in vapA, a homologue of a novel putative transcriptional regulator, out-competed the wild-type strain in the same assay. The virulent phenotype was restored for a mutant strain in orf859 upon complementation. This data provided new insight into the pathogenic personality of A. actinomycetemcomitans in vivo and supported the use of HeLa cells as a valid in vitro host–pathogen interactions model for that microorganism. IVIAT is applicable to most pathogens and will undoubtedly lead to the discovery of novel therapies, antibiotics and diagnostic tools.

In vivo induced antigenic determinants of Actinobacillus actinomycetemcomitans

Actinobacillus actinomycetemcomitans is a Gram-negative capnophilic rod and the etiological agent of localized aggressive periodontitis. The genome-wide survey of A. actinomycetemcomitans using in vivo induced antigen technology (IVIAT) has previously resulted in the discovery of antigenic determinants expressed specifically in diseased patients. The present study evaluated the potential of these antigens as putative disease markers, and investigating their contribution to the pathogenesis of the microorganism. Sera from patients had a significantly greater antibody titer than sera from healthy controls against six antigens, which supports the in vivo expression of these antigens, and suggests their usefulness as disease markers. A. actinomycetemcomitans invasion of epithelium-derived HeLa cells resulted in the induction of all three genes tested, as evidenced by real-time PCR. Isogenic mutants of these three genes were constructed and the adhesion and intracellular survival of the mutants was assayed in a competition assay with the wild-type strain. A significant defect in the intracellular survival of two of these mutant strains ( orf1402 and orf859) was found. This defect could not be attributed to an adhesion defect. In contrast, a mutation in vapA, a homologue of a novel putative transcriptional regulator, out-competed the wild-type strain in the same assay. The virulent phenotype was restored for a mutant strain in orf859 upon complementation. This data provided new insight into the pathogenic personality of A. actinomycetemcomitans in vivo and supported the use of HeLa cells as a valid in vitro host–pathogen interactions model for that microorganism. IVIAT is applicable to most pathogens and will undoubtedly lead to the discovery of novel therapies, antibiotics and diagnostic tools.

Asbestos induces mitochondrial DNA damage and dysfunction linked to the development of apoptosis.

To test the hypothesis that asbestos-mediated cell injury is mediated through an oxidant-dependent mitochondrial pathway, isolated mesothelial cells were examined for mitochondrial DNA damage as determined by quantitative PCR. Mitochondrial DNA damage occurred at fourfold lower concentrations of crocidolite asbestos compared with concentrations required for nuclear DNA damage. DNA damage by asbestos was preceded by oxidant stress as shown by confocal scanning laser microscopy using MitoTracker Green FM and the oxidant probe Redox Sensor Red CC-1. These events were associated with dose-related decreases in steady-state mRNA levels of cytochrome c oxidase, subunit 3 (COIII), and NADH dehydrogenase 5. Subsequently, dose-dependent decreases in formazan production, an indication of mitochondrial dysfunction, increased mRNA expression of pro- and antiapoptotic genes, and increased numbers of apoptotic cells were observed in asbestos-exposed mesothelial cells. The possible contribution of mitochondrial-derived pathways to asbestos-induced apoptosis was confirmed by its significant reduction after pretreatment of cells with a caspase-9 inhibitor. Apoptosis was decreased in the presence of catalase. Last, use of HeLa cells transfected with a mitochondrial transport sequence targeting the human DNA repair enzyme 8-oxoguanine DNA glycosylase to mitochondria demonstrated that asbestos-induced apoptosis was ameliorated with increased cell survival. Studies collectively indicate that mitochondria are initial targets of asbestos-induced DNA damage and apoptosis via an oxidant-related mechanism.

Development of an immunofluorescence method for the detection of antibodies to Ebola virus subtype Reston by the use of recombinant nucleoprotein-expressing HeLa cells.

An indirect immunofluorescent assay (IFA) to detect Ebola virus subtype Reston (EBO-R) antibodies was developed by the use of a HeLa cell line stably expressing EBO-R nucleoprotein (NP). This IFA has a high specificity for the detection of EBO-R IgG antibodies in both hyperimmune rabbit sera and monkey sera collected during an EBO-R outbreak in the Philippines in 1996. Furthermore, this IFA showed a higher sensitivity for the detection of EBO-R antibodies than did the IFA using HeLa cells expressing the NP of Ebola virus subtype Zaire. These results suggest that this new IFA is useful for seroepidemiological studies of EBO-R infection among monkeys.

A Novel High Mobility Group Protein Gene Is a Candidate for Xp22 Abnormalities in Uterine Leiomyomas and Other Benign Tumors

Because genes of the high mobility group protein family HMGI(Y) are known to take part in the development of a variety of benign solid tumors, the aim of the present study was to search for further members of that family in the human genome. Analysis for HMGI(Y)-related sequences by the polymerase chain reaction (PCR) with the use of cDNA-specific primers offered evidence for HMGIY-like sequences, whereas HMGIC-related sequences were apparently absent. By chromosomal assignment of somatic cell hybrids PCR, HMGIY cDNA-related sequences were detected on seven chromosomes. Positive clones were obtained by screening of a P1-derived artificial chromosome library and mapped by fluorescence in situ hybridization. One of these clones assigned to Xp22.1 was chosen for further analysis because Xp22 is a target region for clonal aberrations in benign solid tumors. Sequence analysis of a DNA fragment of this clone, designated as HMGIYL1, revealed a 94.4% homology to the coding region of HMGIY. Within the HMGIYL1 sequence, no nucleotide sequence divergences leading to a frame shift or a new termination codon compared to HMGIY were found, and a TATA-box-like motif 5′ of it was detected. By reverse transcriptase PCR experiments with the use of HeLa cells and human fetal tissue, HMGIYL1 expression was not detectable. Nevertheless, if not active by itself, it is possible that HMGIYL1 may become activated by chromosomal rearrangements of Xp22 observed in benign solid tumors.

The value of LYM-1 cells for examining vacuole formation and loss of cell viability induced by culture supernates of Helicobacter pylori

Some strains of Helicobacter pylori are known to produce an extracellular cytotoxin that causes vacuolation in cultured mammalian cells. Screening for such strains makes use of HeLa cells which may not be sensitive enough to detect minimal changes. The aim of this study was to develop a more sensitive cell line. Vacuole formation was examined in HeLa cells, as well as four other cell lines established in this laboratory by ammonium chloride induction. Among five cell lines tested, LYM-1 cells were most sensitive for the detection of intracellular vacuolation with this agent. Loss of cell viability of LYM-1 and HeLa cells induced by H. pylori culture supernates was also examined: LYM-1 were more sensitive than HeLa cells. Cell death was not always accompanied by vacuole formation. This suggests that the mechanism whereby cell death occurs must be different from that for vacuole formation. LYM-1 cells may be useful when measuring vacuole formation and cell death of the cultured cells induced by culture supernates of clinical isolates of H. pylori.

Old issues confront biotechnology and new products: John Petricciani discusses the risks and rewards associated with the use of abnormal cells for biological production

Few decisions have had such a far-reaching effect on the development and production of biological products as that made in 1954 by the Armed Forces Epidemiological Board. It was at that point that a decision had to be made regarding the use of HeLa cells as the substrate for an experimental adenovirus vaccine. Because HeLa cells were rejected in favour of ‘normal cells’, a philosophy emerged which equated anything other than primary cell cultures from normal tissues of normal animals as dangerous and unacceptable. It was a philosophy which probably made sense at the time, and which had the general support of the biomedical community. However, once it became established as a standard of practice, any suggestion to depart from that position was met with significant obstacles as exemplified by the diploid cell line saga.

Use of HeLa cells infected with tubercle bacilli for the study of antituberculous drugs.

It has been reported elsewhere that the growth of tubercle bacilli in HeLa cells in monolayer is rapid enough to be readily apparent in a few days (Shepard, 1955). T'he present paper describes a method for the employment of this convenient tissue culture system for the study of anti-tuberculous drugs. Mackaness (1952), AMackaness and Smith (1952), and Suter (1952) have used rabbit and guinea pig macrophages in vitro to measure the activity of anti-tuberculous drugs and have pointed out that some of these drugs are not fully active against intracellular bacilli. Thus, the minimal inhibitory concentration of streptomycin in the macrophage system was 10 to 100 times more than it was in the usual test in bacteriological media, whereas the minimal inhibitory concentration of isoniazid, isonicotinic acid hydrazide (INH) was the same in the macrophage system as in bacteriological media. This difference between streptomycin and INH was thought to arise from a relative impermeability of the cell membrane to streptomycin, so that the drug had to be present in excessive concentration in the extracellular fluid in order to achieve an adequate level within the cell. In contrast, the cell membrane was thought to be freely permeable to INH so that it was present in the same concentration within the cell as in the surrounding fluid. This conception took origin in the work of Magoffin and Spink (1951) who showed that brucella organisms were able to escape the action of streptomycin in bacteriological media if they were contained in human leucocytes. The factor of apparent permeability of the cell to the drug was taken into account in the experimental arrangement described below by the adoption of two alternative schedules for the administration of the drug. The two schedules were based on the observation that in this system the phago-

The use of HeLa cells in suspension for the quantitative study of virus propagation

Trypsinized HeLa cells were kept in single-state suspension by means of a magnetic stirrer without a significant loss in concentration. The vast majority of the cells remained in a viable state for at least 24 hours as determined by vital staining. The viability depended to some extent on the initial concentration of the cells. The suspended cells were able to support the propagation of poliomyelitis virus and the development of noninfectious hemagglutinins of influenza A virus, and preliminary quantitative results are presented for these agents.