Cell Based Assays

Telomere “caps” are located at the ends of all chromosomes, including those of the immune system’s T-lymphocytes (T cells), and become shorter with every cell division. Once these chromosomes reach a point where the telomere is too short to provide adequate protection, division ceases and the cell proceeds to senescence. The loss of immune cells to this process negatively impacts the functionality of the immune system, leading to chronic health conditions and/or cancerous diseases. This process is one of the primary factors related to aging, and current dogma suggests that the only way to counteract telomere shortening is through the DNA synthesizing enzyme telomerase. However, an exciting new study published in Nature Cell Biology has identified a previously unknown mechanism of combating telomere aging.

SARS-CoV-2 is the seventh known coronavirus that causes the human disease known as COVID-19. The virus can grow in cells lining the conducting airways and in alveolar epithelial cells. First, the virus generally enters the body through the nose or mouth. From there, the virus travels down into the alveoli which are located in the lungs. Once in the alveoli, the virus “hijacks” cells to make new copies of the virus. The infected cell is then killed, releasing new viruses to infect neighboring cells in the alveolus. Each sac of air, or alveolus, is wrapped with capillaries where red blood cells release carbon dioxide (CO₂) and pick up oxygen (O₂). Two alveolar epithelial cells (type I and II) facilitate gas exchange. Type I cells are squamous alveolar cells with thin membranes that perform gas exchange. Type II cells are known as progenitor cells in the alveoli and proliferate and differentiate into type I cells. In addition, Type II cells secrete the pulmonary surfactant that lines

Traditional 2D cultures have been used widely over the past decades to study cell biology, molecular biology and conduct translation research such as drug discovery. Cells in 2D culture, however, are forced to adopt a planar morphology and maintain cellular interactions only in lateral directions, altering gene transcription, protein translation, and functional phenotypes. As a result, there is a shift towards using 3D in vitro models in the last several years as cell morphology and physiology more closely represent cells in vivo.

There are 2 main types of 3D culture systems known as scaffold-based and scaffold-free. In Table 1 below, the advantages and disadvantages of the different 3D cell culture techniques are listed to help researchers determine the most appropriate 3D culture method for their research.

Table 1: Advantages and Disadvantages of Different 3D Cell Culture Techniques.

Due to their novelties and complexities, 3D cell culture technologies may be daunting to some researchers.

Cell-based assays are widely used in basic and translational research as cost-effective and accessible models to mimic in vivo responses. To obtain reliable data, assessing the health of cultured cells prior to any assays is highly recommended. Furthermore, many cell-based assays require quantification of cell growth. Cell health and growth can be determined by quantifying cell viability, proliferation, or apoptosis.

Below, we compare some commonly used assays to help you determine which type is suitable for your experimental design.

• Cell viability assays enumerate the ratio of live and dead cells in a population. Cell viability can simply be achieved by staining and counting live or dead cells.  A deeper assessment of cell health can be attained by measuring cell metabolic activity, such as the ability to reduce tetrazolium salts in MTT and WST-1 assays.
• Cell proliferation assays assess dividing cells. Some common assays include BrdU incorporation (BrdU Assays)

Primary cells, which are isolated directly from tissue, show normal cell morphology and
maintain many of the important markers and functions seen in vivo. Primary cells, though, have a finite lifespan and limited expansion capacity, so it is critical to use low passage primary cells for your research.

Cell culturing is a widely used technique to grow cells outside of their natural environment using artificial environments and controlled conditions that has become indispensable to scientific research.  The applications of cell culturing are innumerable and there are many ways to differentiate types of cell culturing.  For instance, there are cells isolated from normal or disease models, primary or immortalized, adherent or suspension, 2D or 3D.  This blog post will focus on 2D cell culturing versus 3D cell culturing to ostend ScienCell’s new line of 3D culturing kits that model endothelial tube formation.

Traditional 2D culturing generally involves growing adherent cells on a flat plastic surface such as in a T-75 flask or a tissue culturing dish.  Depending on the cell type, sometimes the plastic surface is coated with an extracellular matrix component or biological compound to promote cell attachment such as fibronectin or poly-L-lysine.  In the case of normal primary cells,

Cancer is a collection of over 200 diseases where the only common denominator is rogue cells^1,2.  The ways in which a cell can go rogue is so varied that cancer has its own separate biology where order and normalcy are not readily apparent.  Cancer does not even have to be solid.  Indeed, blood cancers like leukemia and lymphomas account for about 10% of new cancer diagnoses in the US³.  Our understanding of cancer is continually being refined, and in preparation for our visit to the 2017 American Association for Cancer Research (AACR) conference, this blog post will give a brief overview of human history with cancer, highlight some accomplishments in cancer research, and discuss two future directions for cancer therapy research.

Fossilized bones and mummies of ancient Egypt provide some of the earliest evidence of cancer, and the first recorded description of cancer dates back to circa 3000 BC characterizing breast tumors as a disease for which “there is no treatment⁵.”

Cell-based assays are widely used in basic and translational research as cost-effective and accessible models to mimic in vivo responses. To obtain reliable data, assessing the health of cultured cells prior to any assays is essential. Furthermore, many cell-based assays require quantification of cell growth. Cell health and growth can be determined by quantifying cell viability, proliferation, or apoptosis. Below, we compare some commonly used assays to help you determine which type is suitable for your experimental design.

• Cell viability assays enumerate the ratio of live and dead cells in a population. Cell viability can simply be achieved by staining and counting live or dead cells.  A deeper assessment of cell health can be attained by measuring cell metabolic activity, such as the ability to reduce tetrazolium salts in MTT and WST-1 assays.
• Cell proliferation assays assess dividing cells. Some common assays include BrdU incorporation (BrdU Assays) that can directly measure DNA synthesis,