Figure 1 - available via license: Creative Commons Attribution 4.0 International
Content may be subject to copyright.
Monitoring pH changes in cell culture by phenol red. Phenol red (Phenolsulfonphthalein) is a triphenylmethane dye used in cell cultures to monitor pH changes. Its color exhibits a gradual transition from yellow to red and fuchsia with increasing pH.
Source publication
The cultivation of cells in a favorable artificial environment has become a versatile tool in cellular and molecular biology. Cultured primary cells and continuous cell lines are indispensable in investigations of basic, biomedical, and translation research. However, despite their important role, cell lines are frequently misidentified or contamina...
Contexts in source publication
Context 1
... red also known as phenolsulfonphthalein is the most frequent pH indicator in cell cultures. This water-soluble dye is a yellow zwitterion at low pH, while it changes to a red-colored anion or a fuchsia-colored di-anion at more basic conditions ( Figure 1). Therefore, this dye has been used as an inert pH indicator dye in many tissue culture media to detect pH shifts, waste products of dying cells, or overgrowth of contaminants that typically cause an acidification of the medium. ...
Context 2
... when comparing the 19 mouse STR markers between three different mouse cell lines, each cell line has a unique constellation that allows the unequivocal discrimination of each cell line from the others ( Table 5). 1 (MCA-8-1) 16 14, 15 16 STR 9-2 (MCA-9-2) ND 15 18 STR 11-2 (MCA-11-2) 16 18 16 STR 12-1 (MCA-11-2) 16 19 17 STR 13-1 (MCA-13-1) 17 15 17 STR 15-3 (MCA-15-3) 25.3 21.3 22.3 STR 17-2 (MCA-17-2) 16 13, 15 16 STR 18-3 (MCA-18-3) 16 21 16 STR 19-2 (MCA-19-2) 12 13 13 STR X-1 (MCA-X-1) 26, 27 26 27 * Representative STR electropherograms of selected markers (i.e., STR 17-2, STR 18-3, STR 19-2, and STR X-1) of this cell line are depicted in Figure 6. ND, not determined. ...
Citations
... risks the validity of research [5,6]. Addressing this issue is essential for ensuring reliable scientific outcomes. ...
... However, even with authentication, cell lines may not accurately represent their original tissues or diseases due to factors such as source and reagent differences [78,79]. Therefore, the validity of a cell line as a research model, not only in the field of NPCs, should always be continually improved [80,81]. ...
Nasopharyngeal carcinoma (NPC) is a type of cancer that originates from the mucosal lining of the nasopharynx and can invade and spread. Although contemporary chemoradiotherapy effectively manages the disease locally, there are still challenges with locoregional recurrence and distant failure. Therefore, it is crucial to have a deeper understanding of the molecular basis of NPC cell movement in order to develop a more effective treatment and to improve patient survival rates. Cancer cell line models are invaluable in studying health and disease and it is not surprising that they play a critical role in NPC research. Consequently, scientists have established around 80 immortalized human NPC lines that are commonly used as in vitro models. However, over the years, it has been observed that many cell lines are misidentified or contaminated by other cells. This cross-contamination leads to the creation of false cell lines that no longer match the original donor. In this commentary, we discuss the impact of misidentified NPC cell lines on the scientific literature. We found 1159 articles from 2000 to 2023 that used NPC cell lines contaminated with HeLa cells. Alarmingly, the number of publications and citations using these contaminated cell lines continued to increase, even after information about the contamination was officially published. These articles were most commonly published in the fields of oncology, pharmacology, and experimental medicine research. These findings highlight the importance of science policy and support the need for journals to require authentication testing before publication.
... The simplest and most economical way to provide aseptic conditions is to use a cell culture hood (i.e., biosafety cabinet). The GCCP guidelines provide background on culture systems, documentation, general safety instructions, training information, also focus on quality management and ethical issues associated with the performance of cell culture experiments [1,2]. ...
Cultivation of animal cell in culture is an utmost requirement of molecular and biomedical research in today’s scenario. All the scientists and researchers conducting research on human genetics, diseases, drug discovery and translational biology are in crucial need to have done invitro investigations. Continuous and primary cell lines need to be established with their authorized details and most importantly without any contamination, under aseptic conditions. In addition, all the manipulations related to cell culture needs proper biosafety cabinets and safeguard procedures to minimize the risk of any contamination and exposure towards any hazardous chemical or biological product. This draft provides a brief introduction to cell culture and guidelines related to culture techniques and maintaining cell lines, along with some safety and precautionary measures to prevent and tackle problems associated with basic cell culture laboratory.
Keywords: Cell culture; Cell lines manipulation; Safety protocols; Culture conditions; Viability assay
... This system also provides deeper insights into metabolic activities, tracking changes in pH and ion concentrations, like acid production during fermentation [37]. It allows the earlier detection of contamination or anomalies, as pH and sodium ion fluctuations indicate such issues more promptly than absorbance alone [38,39]. In microgravity environments like those in CubeSats, these detailed data are essential for understanding yeast behavior in space, vital for long-term space missions. ...
CubeSats have emerged as cost-effective platforms for biological research in low Earth orbit (LEO). However, they have traditionally been limited to optical absorbance sensors for studying microbial growth. This work has made improvements to the sensing capabilities of these small satellites by incorporating electrochemical ion-selective pH and pNa sensors with optical absorbance sensors to enrich biological experimentation and greatly expand the capabilities of these payloads. We have designed, built, and tested a multi-modal multi-array electrochemical-optical sensor module and its ancillary systems, including a fluidic card and an on-board payload computer with custom firmware. Laboratory tests showed that the module could endure high flow rates (1 mL/min) without leakage, and the 27-well, 81-electrode sensor card accurately detected pH (71.0 mV/pH), sodium ion concentration (75.2 mV/pNa), and absorbance (0.067 AU), with the sensors demonstrating precise linear responses (R2 ≈ 0.99) in various test solutions. The successful development and integration of this technology conclude that CubeSat bio-payloads are now poised for more complex and detailed investigations of biological phenomena in space, marking a significant enhancement of small-satellite research capabilities.
... Fibroblastic lines are defined as populations of fibroblasts that proliferate in vitro over an extended period when given an appropriate medium and space, and therefore can be maintained by serial subculturing while maintaining their morphological and physiological characteristics [1]. Although they represent part of an animal's tissue, they can be used for extensive in vitro experimentations to understand cell behavior through analyzes such as morphology evaluation, population doubling time, metabolism performance, and tolerance to oxidative stress [2]. ...
The loss of wild biodiversity has encouraged the development of fibroblastic lines, mainly fibroblasts derived from skin, which can be interesting tools for the conservation of wild mammals. These biological samples, when properly well-established, are essential elements for the reproduction of species through their use in cloning by somatic cell nuclear transfer and induction of cells to pluripotency. In general, the establishment of fibroblastic lines involves the following strategies: (i) cell isolation techniques and identification of fibroblasts; (ii) conditions for in vitro culture of fibroblasts; (iii) conditions for cryopreservation of fibroblasts; and (iv) nuclear reprogramming studies. At each stage, species-specific factors are involved, and determining these lines in the species of interest represents the first step toward its successful use for animal conservation. Therefore, this chapter discusses the stages and parameters involved in the strategies for establishing fibroblastic lines, delving into the main technical aspects and results obtained from the use of these cells in recent years in wild mammals.
... Unfortunately, the identification of a misidentified cell line is complicated because cell lines often have several synonyms, just like in the case of L-O2 or SMMC-7721. In addition, workflows for quality control and cell line authentication testing should be established in laboratories working with established cell lines (24). In particular, cell line identity testing using short tandem repeat (STR) profiling is an effective means to prevent cell line misidentification and to identify cross-contamination at early stage (24). ...
... In addition, workflows for quality control and cell line authentication testing should be established in laboratories working with established cell lines (24). In particular, cell line identity testing using short tandem repeat (STR) profiling is an effective means to prevent cell line misidentification and to identify cross-contamination at early stage (24). Guidelines and guidance for proper documentation of experiments that are conducted with continuous cell lines are published elsewhere (25). ...
... Electrophoresis is an analytical and preparative method for separating electrically charged particles and particle aggregates under the influence of a uniformly applied external electric field. The technique relies on the physicochemical phenomenon of differential movement or migration of different particles within an electric field (60). Morphologically, no significant differences exist between the analyzed cases and the control. ...
Primary fibroblast cultures obtained from Wistar rats were investigated with a focus on two vital physiological mechanisms: inflammatory processes and oxidative stress balance. These are believed to be affected by mud and sulfurous natural mineral waters, forming the fundamental biological basis for understanding the therapeutic effects of these substances. Existing scientific research highlights that various cell types, including fibroblasts, are recruited during inflammation. These cells respond to a wide array of intercellular and microenvironmental signals, leading to a regulated production of both pro- and anti-inflammatory mediators. Examples include cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, as well as chemokines and enzymes like cyclooxygenase (COX)-2. Together, these play vital roles in modulating the inflammatory response.
... For this reason, it is essential that cell cultures are kept aseptic. Although different microorganisms (bacterial, fungi, yeast, or viruses) are capable of contaminating cell cultures [4], Mollicutes (or interchangeably Mycoplasma) are the fundamental cause of contamination. Despite the availability of sophisticated technology and specialisations used in manipulating cell cultures, biological contamination primarily due to Mycoplasma contamination remains a significant challenge. ...
In recent years, cell culture has become an important tool not only in research laboratories, but also in diagnostic and biotechnological development laboratories. Mycoplasma contamination is present in up to 35% of cell cultures used in research and in cell therapies. This fact represents a significant problem since such contamination can cause disastrous effects on eukaryotic cells by altering their cellular parameters, which, in turn, can lead to unreliable experimental results. For this reason, it is mandatory to carry out continuous testing for the presence of Mycoplasma in cell culture and the development of appropriate methodologies for this purpose. An ideal detection methodology should be fast, sensitive, and reliable. In this study, we propose an alternative detection method based on real-time PCR in conjunction with a novel combination of primers and probes that have been improved to increase their efficiency. The new PCR method demonstrates 100% sensitivity and specificity results in the detection of common Mycoplasma species that contaminate cell cultures. Whilst 11 of 45 tested supernatants were positive for Mycoplasma (24.4%) using the new PCR method (corresponding to 5 of the 14 lines tested (35.71%)), only 10 of 45 supernatants showed positive results with the commercial Venor®GeM qEP and Plasmotest® kit. In addition, the new PCR method exhibits a high capacity to detect less-frequent Mycoplasma species, such as those related to the M. mycoides cluster. The use of an alternative Mycoplasma-detection method in cell culture labs can guarantee the detection of Mycoplasma contamination, especially in cases when dubious results are recorded.
... The elimination of mycoplasma in PAV-1 culture enables the safe application of this cell line in many laboratories and deposition in suitable cell line banks. Nevertheless, as recommended for all other established cell lines, regular testing for inter-and intra-specific cross-contamination and bacterial impurities is still highly recommended to prevent falsified research results, misleading publications, and waste of research money [68]. ...
The rat hepatic stellate cell line PAV-1 was established two decades ago and proposed as a cellular model to study aspects of hepatic retinoic acid metabolism. This cell line exhibits a myofibroblast-like phenotype but also has the ability to store retinyl esters and synthesize retinoic acid from its precursor retinol. Importantly, when cultured with palmitic acid alone or in combination with retinol, the cells switch to a deactivated phenotype in which the proliferation and expression of profibrogenic marker genes are suppressed. Despite these interesting characteristics, the cell line has somehow fallen into oblivion. However, based on the fact that working with in vivo models is becoming increasingly complicated, genetically characterized established cell lines that mimic aspects of hepatic stellate cell biology are of fundamental value for biomedical research. To genetically characterize PAV-1 cells, we performed karyotype analysis using conventional chromosome analysis and multicolor spectral karyotyping (SKY), which allowed us to identify numerical and specific chromosomal alteration in PAV-1 cells. In addition, we used a panel of 31 species-specific allelic variant sites to define a unique short tandem repeat (STR) profile for this cell line and performed bulk mRNA-sequencing, showing that PAV-1 cells express an abundance of genes specific for the proposed myofibroblastic phenotype. Finally, we used Rhodamine-Phalloidin staining and electron microscopy analysis, which showed that PAV-1 cells contain a robust intracellular network of filamentous actin and process typical ultrastructural features of hepatic stellate cells.
