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Cell Death: The termination of the cell's ability to carry out vital functions such as metabolism, growth, reproduction, responsiveness, and adaptability.

An Introduction to Cell Death

JoVE 5649

Necrosis, apoptosis, and autophagic cell death are all manners in which cells can die, and these mechanisms can be induced by different stimuli, such as cell injury, low nutrient levels, or signaling proteins. Whereas necrosis is considered to be an “accidental” or unexpected form of cell death, evidence exists that apoptosis and autophagy are both programmed…

 Cell Biology

Assay for Cell Death: Chromium Release Assay of Cytotoxic Ability

JoVE 10505

Source: Frances V. Sjaastad1,2, Whitney Swanson2,3, and Thomas S. Griffith1,2,3,4
1 Microbiology, Immunology, and Cancer Biology Graduate Program, University of Minnesota, Minneapolis, MN 55455
2 Center for Immunology, University of Minnesota, Minneapolis, MN 55455
3 Department of Urology, University of Minnesota, Minneapolis, MN 55455
4 Masonic…


Cell Division- Concept

JoVE 10571

Cell division is fundamental to all living organisms and required for growth and development. As an essential means of reproduction for all living things, cell division allows organisms to transfer their genetic material to their offspring. For a unicellular organism, cellular division generates a completely new organism. For multicellular organisms, cellular division produces new cells for…

 Lab Bio

What is the Cell Cycle?

JoVE 10757

The cell cycle refers to the sequence of events occurring throughout a typical cell’s life. In eukaryotic cells, the somatic cell cycle has two stages: interphase and the mitotic phase. During interphase, the cell grows, performs its basic metabolic functions, copies its DNA, and prepares for mitotic cell division. Then, during mitosis and cytokinesis, the cell divides its nuclear and cytoplasmic materials, respectively. This generates two daughter cells that are identical to the original parent cell. The cell cycle is essential for the growth of the organism, replacement of damaged cells, and regeneration of aged cells. Cancer is the result of uncontrolled cell division sparked by a gene mutation. There are three major checkpoints in the eukaryotic cell cycle. At each checkpoint, the progression to the next cell cycle stage can be halted until conditions are more favorable. The G1 checkpoint is the first of these, where a cell’s size, energy, nutrients, DNA quality, and other external factors are evaluated. If the cell is deemed inadequate, it does not continue to the S phase of interphase. The G2 checkpoint is the second checkpoint. Here, the cell ensures that all of the DNA has been replicated and is not damaged before entering mitosis. If any DNA damage is detected that cannot be repaired, the cell may undergo apoptosis, or

 Core: Cell Cycle and Division

An Introduction to Cell Division

JoVE 5640

Cell division is the process by which a parent cell divides and gives rise to two or more daughter cells. It is a means of reproduction for single-cell organisms. In multicellular organisms, cell division contributes to growth, development, repair, and the generation of reproductive cells (sperms and eggs). Cell division is a tightly regulated process, and aberrant cell…

 Cell Biology

Live Cell Imaging of Mitosis

JoVE 5642

Mitosis is a form of cell division in which a cell’s genetic material is divided equally between two daughter cells. Mitosis can be broken down into six phases, during each of which the cell’s components, such as its chromosomes, show visually distinct characteristics. Advances in fluorescence live cell imaging have allowed scientists to study this process in…

 Cell Biology

Negative Regulator Molecules

JoVE 10764

Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.

Three of the best-understood negative regulators are p53, p21, and retinoblastoma protein (Rb). The regulatory roles of each of these proteins were discovered after faulty copies were found in cells with uncontrolled replication (i.e., cancer). These proteins exert most of their regulatory effects at the G1 checkpoint early in the cell cycle. P53 strongly influences a cell’s commitment to divide. It responds to DNA damage by discontinuing the cell cycle and summoning enzymes to repair the damage. If the DNA damage is irreparable, p53 can prevent the cell from proceeding through the cell cycle by inducing apoptosis, or cell death. An increase in p53 triggers the production of p21. P21 prevents the cell from transitioning from the G1 to the S phase of the cell cycle by binding to CDK/cyclin complexes, inhibiting their positive regulatory actions. Rb negatively regulates the cell cycle by acting on different positive regulators, mainly in response to cell size. Active (dephosphorylated) Rb binds to transcription factors, preventing them from initiating gene tran

 Core: Cell Cycle and Division

Annexin V and Propidium Iodide Labeling

JoVE 5650

Staining with annexin V and propidium iodide (PI) provides researchers with a way to identify different types of cell death—either necrosis or apoptosis. This technique relies on two components. The first, annexin V, is a protein that binds certain phospholipids called phosphatidylserines, which normally occur only in the inner, cytoplasm-facing leaflet of a…

 Cell Biology

Yeast Signaling

JoVE 10714

Yeasts are single-celled organisms, but unlike bacteria, they are eukaryotes—cells that have a nucleus. Cell signaling in yeast is similar to signaling in other eukaryotic cells. A ligand, such as a protein or a small molecule outside the yeast cell, attaches to a receptor on the cell surface. The binding stimulates second-messenger kinases (enzymes that phosphorylate specific substrates) to activate or inactivate transcription factors that regulate gene expression. Many of the yeast intracellular signaling cascades have similar counterparts in Homo sapiens, making yeast a convenient model for studying intracellular signaling in humans. Yeasts are members of the fungus kingdom. They use signaling for various functions, especially for reproduction. Yeasts can undergo “sexual” reproduction using mating pheromones, which are peptides—short chains of amino acids. Yeast colonies consist of both diploid and haploid cells. Both types of cells can undergo mitosis, but only diploid cells can undergo meiosis. When diploid cells undergo meiosis, the four resulting haploid cells, called spores, are not identical. In fact, the division of one diploid cell into four spores creates two “sexes” of yeast cells, each two cells of the type MAT-a and MAT-alpha. MAT-a cells secrete mating

 Core: Cell Signaling

Identification of Intracellular Signaling Events Induced in Viable Cells by Interaction with Neighboring Cells Undergoing Apoptotic Cell Death

1Section of Nephrology, Department of Medicine, University of Illinois at Chicago, 2Section of Nephrology, Department of Medicine, Jesse Brown Veterans Affairs Medical Center, 3Department of Biology, Kutztown University of Pennsylvania, 4Division of Rheumatology, Department of Medicine, Research Institute of the McGill University Health Centre, 5Department of Microbiology & Immunology, University of Illinois at Chicago

JoVE 54980


Adult Stem Cells

JoVE 10810

Stem cells are undifferentiated cells that divide and produce more stem cells or progenitor cells that differentiate into mature, specialized cell types. All the cells in the body are generated from stem cells in the early embryo, but small populations of stem cells are also present in many adult tissues including the bone marrow, brain, skin, and gut. These adult stem cells typically produce the various cell types found in that tissue—to replace cells that are damaged or to continuously renew the tissue. The epithelium lining the small intestine is continuously renewed by adult stem cells. It is the most rapidly replaced tissue in the human body, with most cells being replaced within 3-5 days. The intestinal epithelium consists of thousands of villi that protrude into the interior of the small intestine—increasing its surface area to aid in the absorption of nutrients. Intestinal stem cells are located at the base of invaginations called crypts that lie between the villi. They divide to produce new stem cells, as well as daughter cells (called transit amplifying cells) that divide rapidly, move up the villi and differentiate into all the cell types in the intestinal epithelium, including absorptive, goblet, enteroendocrine, and Paneth cells. These mature cells continue to move up the villi as they carry out their functions, except Paneth cell

 Core: Biotechnology

An Introduction to Saccharomyces cerevisiae

JoVE 5081

Saccharomyces cerevisiae (commonly known as baker’s yeast) is a single-celled eukaryote that is frequently used in scientific research. S. cerevisiae is an attractive model organism due to the fact that its genome has been sequenced, its genetics are easily manipulated, and it is very easy to maintain in the lab. Because many yeast proteins are similar in sequence and function…

 Biology I

Yeast Maintenance

JoVE 5095

Research performed in the yeast Saccharomyces cerevisiae has significantly improved our understanding of important cellular phenomona such as regulation of the cell cycle, aging, and cell death. The many benefits of working with S. cerevisiae include the facts that they are inexpensive to grow in the lab and that many ready-to-use strains are now commercially available. Nevertheless,…

 Biology I

RNA Splicing

JoVE 10802

The process in which eukaryotic RNA is edited prior to protein translation is called splicing. It removes regions that do not code for proteins and patches the protein-coding regions together. Splicing also allows several protein variants to be expressed from a single gene and plays an essential role in development, tissue differentiation, and adaptation to environmental stress. Errors in splicing can lead to diseases such as cancer. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts designated to become mRNA are called precursor messenger RNA (pre-mRNA). The pre-mRNA is then processed to form mature mRNA that is suitable for protein translation. Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins whereas introns are the non-coding regions. RNA splicing is the process by which introns are removed and exons patched together. Splicing is mediated by the spliceosome—a complex of proteins and RNA called small nuclear ribonucleoproteins (snRNPs). The spliceosome recognizes specific nucleotide sequences at exon/intron boundaries. First, it binds to a GU-containing sequence at the 5’ end of the intron and to a branch point sequence containing an A towards the 3’ end of the intron. In a number of carefully-orches

 Core: Gene Expression

The TUNEL Assay

JoVE 5651

One of the hallmarks of apoptosis is the nuclear DNA fragmentation by nucleases. These enzymes are activated by caspases, the family of proteins that execute the cell death program. TUNEL assay is a method that takes advantage of this feature to detect apoptotic cells. In this assay, an enzyme called terminal deoxynucleotidyl transferase catalyzes the addition of dUTP…

 Cell Biology

C. elegans Development and Reproduction

JoVE 5110

Ceanorhabditis elegans is a powerful tool to help understand how organisms develop from a single cell into a vast interconnected array of functioning tissues. Early work in C. elegans traced the complete cell lineage and structure at the electron microscopy level, allowing researchers unprecedented insight into the connection between genes, development and disease. …

 Biology I

Nucleotide Excision Repair

JoVE 10792

Exposure to mutagens can damage DNA and result in bulky lesions that distort the double-helix structure or impede proper transcription. Damaged DNA can be detected and repaired in a process called nucleotide excision repair (NER). NER employs a set of specialized proteins that first scan DNA to detect a damaged region. Next, NER proteins separate the strands and excise the damaged area. Finally, they coordinate the replacement with new, matching nucleotides. Cells are regularly exposed to mutagens—factors in the environment which can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes to DNA. These include bends or kinks in the structure which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations which in turn can result in cancer or disease depending on which sequences are disrupted. Nucleotide excision repair relies on specific protein complexes to recognize damaged regions of DNA and flag them for removal and repair. In prokaryotes, the process involves three proteins—UvrA, UvrB, and UvrC. The first two proteins work together as a complex, traveling along the DNA strands to detect any physical aberrations. Once identified, the strands at the damaged location are separated, and endon

 Core: DNA Structure and Function

Bacterial Growth Curve Analysis and its Environmental Applications

JoVE 10100

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - Arizona University
Demonstrating Author: Luisa Ikner

Bacteria are among the most abundant life forms on Earth. They are found in every ecosystem and are vital for everyday life. For example, bacteria affect what people eat, drink, and breathe, and there are actually more…

 Environmental Microbiology

Histological Sample Preparation for Light Microscopy

JoVE 5039

Histology is the study of cells and tissues, which is typically aided by the use of a light microscope. The preparation of histological samples can vary greatly based on the inherent properties of the samples such as size and hardness as well as expected post-processing which includes planned staining techniques or other down-stream applications. As described in this video, specimen…

 General Laboratory Techniques

An Introduction to Caenorhabditis elegans

JoVE 5103

Caenorhabditis elegans is a microscopic, soil-dwelling roundworm that has been powerfully used as a model organism since the early 1970’s. It was initially proposed as a model for developmental biology because of its invariant body plan, ease of genetic manipulation and low cost of maintenance. Since then C. elegans has rapidly grown in popularity and is now utilized…

 Biology I

Fundamentals of Breeding and Weaning

JoVE 10293

Source: Kay Stewart, RVT, RLATG, CMAR; Valerie A. Schroeder, RVT, RLATG. University of Notre Dame, IN

Millions of mice and rats are bred for use in biomedical research each year. Worldwide, there are several large commercial breeding facilities that supply mice to research laboratories, but many facilities choose to also breed mice and…

 Lab Animal Research

Use of Single Chain MHC Technology to Investigate Co-agonism in Human CD8+ T Cell Activation

1Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 2Immunology Programme, Life Sciences Institute, National University of Singapore, 3Emerging Infectious Diseases Program, Duke-NUS Graduate Medical School, 4Singapore Immunology Network, A*STAR, 5NUS Graduate School for Integrative Sciences and Engineering (NGS), National University of Singapore, 6Department of Immunology, Wright-Fleming Institute, Imperial College London

JoVE 59126

 Immunology and Infection

Primary Cell Culture of Purified GABAergic or Glutamatergic Neurons Established through Fluorescence-activated Cell Sorting

1Institut für Integrative Neuroanatomie, Charité, Universitätsmedizin Berlin, 2Department for Cytometry and Cellsorting, German Rheumatism Research Center, Leibniz Institute, 3Departments of Genetic and Behavioral Neuroscience, Graduate School of Medicine, Gunma University

JoVE 58974


Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues

1Hotchkiss Brain Institute, University of Calgary, 2Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, 3Alberta Children's Hospital Research Institute, University of Calgary, 4Department of Clinical Neurosciences, Cumming School of Medicine, University of Calgary

JoVE 58709


Flow Cytometry-based Assay for the Monitoring of NK Cell Functions

1Childrens Hospital, Department of Pediatric Stem Cell Transplantation and Immunology, Johann Wolfgang Goethe-University, 2LOEWE Center for Cell and Gene Therapy, Johann Wolfgang Goethe-University, 3Institute for Cancer Research, Department of Cancer Immunology, Oslo University Hospital, Radiumhospital, 4The KG Jebsen Center for Cancer Immunotherapy, Institute of Clinical Medicine, University of Oslo

JoVE 54615

 Immunology and Infection

Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids

1The Centenary Institute, 2Sydney Medical School, University of Sydney, 3The University of Queensland Diamantina Institute, Translational Research Institute, The University of Queensland, 4Department of Dermatology, Royal Prince Alfred Hospital, 5Discipline of Dermatology, University of Sydney

JoVE 53486


Construction of Defined Human Engineered Cardiac Tissues to Study Mechanisms of Cardiac Cell Therapy

1Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, 2The Mindich Child Health and Development Institute, Icahn School of Medicine at Mount Sinai, 3Stem Cell & Regenerative Medicine Consortium, LKS Faculty of Medicine, University of Hong Kong

JoVE 53447


Time-lapse Imaging of Primary Preneoplastic Mammary Epithelial Cells Derived from Genetically Engineered Mouse Models of Breast Cancer

1Department of Oncology, Georgetown University, 2Lombardi Comprehensive Cancer Center, Georgetown University, 3Stem Cell Dynamics, Helmholtz Zentrum München - German Research Center for Environmental Health, 4Department of Medicine, Georgetown University, 5Department of Nanobiomedical Science and WCU Research Center of Nanobiomedical Science, Dankook University

JoVE 50198

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