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Journal of Hepatology Oct 2014The liver has the unique capacity to regenerate in response to a damaging event. Liver regeneration is hereby largely driven by hepatocyte proliferation, which in turn... (Review)
Review
The liver has the unique capacity to regenerate in response to a damaging event. Liver regeneration is hereby largely driven by hepatocyte proliferation, which in turn relies on cell cycling. The hepatocyte cell cycle is a complex process that is tightly regulated by several well-established mechanisms. In vitro, isolated hepatocytes do not longer retain this proliferative capacity. However, in vitro cell growth can be boosted by immortalization of hepatocytes. Well-defined immortalization genes can be artificially overexpressed in hepatocytes or the cells can be conditionally immortalized leading to controlled cell proliferation. This paper discusses the current immortalization techniques and provides a state-of-the-art overview of the actually available immortalized hepatocyte-derived cell lines and their applications.
Topics: Animals; Cell Culture Techniques; Cell Proliferation; Cells, Cultured; Cellular Senescence; Hepatocytes; Humans
PubMed: 24911463
DOI: 10.1016/j.jhep.2014.05.046 -
Cell Feb 2008The cell-cycle transition from G1 to S phase has been difficult to visualize. We have harnessed antiphase oscillating proteins that mark cell-cycle transitions in order...
The cell-cycle transition from G1 to S phase has been difficult to visualize. We have harnessed antiphase oscillating proteins that mark cell-cycle transitions in order to develop genetically encoded fluorescent probes for this purpose. These probes effectively label individual G1 phase nuclei red and those in S/G2/M phases green. We were able to generate cultured cells and transgenic mice constitutively expressing the cell-cycle probes, in which every cell nucleus exhibits either red or green fluorescence. We performed time-lapse imaging to explore the spatiotemporal patterns of cell-cycle dynamics during the epithelial-mesenchymal transition of cultured cells, the migration and differentiation of neural progenitors in brain slices, and the development of tumors across blood vessels in live mice. These mice and cell lines will serve as model systems permitting unprecedented spatial and temporal resolution to help us better understand how the cell cycle is coordinated with various biological events.
Topics: Animals; COS Cells; Cell Cycle; Cell Cycle Proteins; Cell Line; Cells, Cultured; Chlorocebus aethiops; Cytological Techniques; Fluorescence; Geminin; HeLa Cells; Humans; Luminescent Proteins; Mice; Mice, Transgenic; Microscopy, Confocal; Molecular Sequence Data; Morphogenesis; Neoplasms; Recombinant Fusion Proteins; Ubiquitination
PubMed: 18267078
DOI: 10.1016/j.cell.2007.12.033 -
Cells Feb 2023The cultivation of cells in a favorable artificial environment has become a versatile tool in cellular and molecular biology. Cultured primary cells and continuous cell... (Review)
Review
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 contaminated by other cells, bacteria, fungi, yeast, viruses, or chemicals. In addition, handling and manipulating of cells is associated with specific biological and chemical hazards requiring special safeguards such as biosafety cabinets, enclosed containers, and other specialized protective equipment to minimize the risk of exposure to hazardous materials and to guarantee aseptic work conditions. This review provides a brief introduction about the most common problems encountered in cell culture laboratories and some guidelines on preventing or tackling respective problems.
Topics: Cell Culture Techniques; Cell Line; Cells, Cultured; Containment of Biohazards
PubMed: 36899818
DOI: 10.3390/cells12050682 -
Infection and Immunity May 2023Brucella spp. are facultatively intracellular bacteria that can infect, survive, and multiply in various host cell types and/or . The genus Brucella has markedly... (Review)
Review
Brucella spp. are facultatively intracellular bacteria that can infect, survive, and multiply in various host cell types and/or . The genus Brucella has markedly expanded in recent years with the identification of novel species and hosts, which has revealed additional information about the cell and tissue tropism of these pathogens. Classically, Brucella spp. are considered to have tropism for organs that contain large populations of phagocytes such as lymph nodes, spleen, and liver, as well as for organs of the genital system, including the uterus, epididymis, testis, and placenta. However, experimental infections of several different cultured cell types indicate that Brucella may actually have a broader cell tropism than previously thought. Indeed, recent studies indicate that certain Brucella species in particular hosts may display a pantropic distribution . This review discusses the available knowledge on cell and tissue tropism of Brucella spp. in natural infections of various host species, as well as in experimental animal models and cultured cells.
Topics: Animals; Male; Female; Brucella; Phagocytes; Cell Line; Cells, Cultured; Tropism; Brucellosis
PubMed: 37129522
DOI: 10.1128/iai.00062-23 -
Cell Host & Microbe Apr 2022
Topics: Cells, Cultured
PubMed: 35421329
DOI: 10.1016/j.chom.2022.03.032 -
Cold Spring Harbor Perspectives in... Nov 2019Hepatitis C virus (HCV) infection is seen worldwide and is a significant cause of severe chronic liver diseases. Recently, a large number of direct-acting antivirals... (Review)
Review
Hepatitis C virus (HCV) infection is seen worldwide and is a significant cause of severe chronic liver diseases. Recently, a large number of direct-acting antivirals (DAAs) have been developed against HCV infection, resulting in significant improvements in treatment efficacy. Rapid progress in HCV research has been largely dependent on the development of HCV culture systems and small animal infection models. In the development of HCV cell culture systems, the discovery of the JFH-1 clone, an HCV strain isolated from a fulminant hepatitis C patient, was a key finding. The JFH-1 strain was the first infectious HCV strain belonging to genotype 2a. JFH-1 replicated efficiently in cultured cell lines without acquiring adaptive mutations, providing the secretion of infectious viral particles into the culture medium. Recently, other HCV strains also were reported to be infectious in cultured cells with adaptive viral mutations, but genotype-1b infectious HCV clones and virus culture systems for clinical isolates are still missing. These infectious HCV systems have provided powerful tools to study the viral life cycle, to construct antiviral strategies, and to develop effective vaccines.
Topics: Animals; Cell Culture Techniques; Cell Line, Tumor; Genotype; Hepacivirus; Humans; RNA, Viral; Virus Cultivation; Virus Replication
PubMed: 31501261
DOI: 10.1101/cshperspect.a036806 -
ELife May 2023Changes in gene expression in cultured endothelial cells can be partially reversed by simulating in vivo conditions.
Changes in gene expression in cultured endothelial cells can be partially reversed by simulating in vivo conditions.
Topics: Endothelial Cells; Cells, Cultured; Gene Expression
PubMed: 37158991
DOI: 10.7554/eLife.88248 -
Current Protocols in Cell Biology May 2001This unit provides a protocol for indirect immunofluorescence, which is a method that provides information about the locations of specific molecules and the structure of...
This unit provides a protocol for indirect immunofluorescence, which is a method that provides information about the locations of specific molecules and the structure of the cell. Antibody molecules for a specific target molecule are exposed to the cell or tissue being investigated. The binding of these molecules is detected by incubating the sample with a secondary antibody specific for immunoglobulin molecules and conjugated to fluorophore. This provides both a visible signal and amplification of the signal and the results are observed with a fluorescent microscope.
Topics: Animals; Antigen-Antibody Reactions; Cells, Cultured; Fluorescent Antibody Technique, Indirect; Fluorescent Dyes; Indicators and Reagents; Microscopy, Fluorescence; Specimen Handling; Staining and Labeling; Tissue Fixation
PubMed: 18228363
DOI: 10.1002/0471143030.cb0403s00 -
STAR Protocols Sep 2023Mammalian cells can die by apoptosis or by one of several non-apoptotic mechanisms, such as ferroptosis. Here, we present a protocol to distinguish ferroptosis from...
Mammalian cells can die by apoptosis or by one of several non-apoptotic mechanisms, such as ferroptosis. Here, we present a protocol to distinguish ferroptosis from other cell death mechanisms in cultured cells. We describe steps for seeding cells, administering mechanism-specific cell death inducers and inhibitors, and measuring cell death and viability. We then detail the use of molecular markers to verify mechanisms of cell death. This protocol can be used to identify and distinguish ferroptosis in 2D and 3D cultures. For complete details on the use and execution of this protocol, please refer to Ko, et al. (2019), Magtanong, et al. (2022), and Armenta, et al. (2022)..
Topics: Animals; Ferroptosis; Cell Death; Apoptosis; Cells, Cultured; Mammals
PubMed: 37556320
DOI: 10.1016/j.xpro.2023.102457 -
Biochimica Et Biophysica Acta Jan 2005The two histopathological signatures of Alzheimer's disease (AD) are amyloid plaques and neurofibrillary tangles, prompting speculation that a causal relationship exists... (Review)
Review
The two histopathological signatures of Alzheimer's disease (AD) are amyloid plaques and neurofibrillary tangles, prompting speculation that a causal relationship exists between the respective building blocks of these abnormal brain structures: the beta-amyloid peptides (Abeta) and the neuron-enriched microtubule-associated protein called tau. Transgenic mouse models have provided in vivo evidence for such connections, and cultured cell models have allowed tightly controlled, systematic manipulation of conditions that influence links between Abeta and tau. The emerging evidence supports the view that amyloid pathology lies upstream of tau pathology in a pathway whose details remain largely mysterious. In this communication, we review and discuss published work about the Abeta-tau connection. In addition, we present some of our own previously unpublished data on the effects of exogenous Abeta on primary brain cultures that contain both neurons and glial cells. We report here that continuous exposure to 5 microM non-fibrillar Abeta40 or Abeta42 kills primary brain cells by apoptosis within 2-3 weeks, Abeta42 is more toxic and selective for neurons than Abeta40, and Abeta42, but not Abeta40, induces a transient increase in neurons that are positive for the AD-like PHF1 epitope. These findings demonstrate the greater potency of Abeta42 than Abeta40 at inducing tau pathology and programmed cell death, and corroborate and extend reports that tau-containing cells are more sensitive to Abeta peptides than cells that lack or express low levels of tau.
Topics: Amyloid beta-Peptides; Animals; Apoptosis; Brain; Cells, Cultured; Humans; Mice; Mice, Transgenic; Neurons; Tauopathies; tau Proteins
PubMed: 15615631
DOI: 10.1016/j.bbadis.2004.08.008