Adherent Cell Culture Virtual Lab using Mammalian Cell Lines

Biology | Toxicology | Biochemistry | Proteomics | Pharmacology

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General Aim of Cell Culture Virtual Lab

  •  The cell culture simulation aims at showing how cells are cultured, propagated, subcultured, and frozen in liquid nitrogen for long term storage. 

In Vitro Cell Culturing is a very important protocol for preparing cells needed for all subsequent In Vitro Cytotoxicity and Genotoxicity assays. 

Method of Cell Culture Virtual Lab

In Vitro Cell Culturing using Adherent Mammalian Cell Lines

Learning Objectives (ILOs) of Mammalian Cell Culture Techniques

  •  By the end of cell culture virtual lab, the postgraduate student will be able to:  

  • Successfully handle the required instruments and consumables needed in cell culturing and subculturing.
  • Strictly work and follow the aseptic technique in cell culture virtual lab.
  • Thaw cells from Liquid Nitrogen and seed them in cell culture flasks.
  • Check the confluence, harvest cells, and count them microscopically.
  • Scale up the cultured cells for setting up further experiments.
  • Freeze cells in Liquid Nitrogen for long-term storage.
  • Work and follow the general safety guidance for Good Laboratory Practice (GLP) in the cell culture virtual lab 

     

Theoretical Background / Context of Adherent Cell Culture

  • Cell culture is the growth of cells from an animal in an artificial, controlled environment. Cells are removed either from the organism directly and disaggregated before cultivation or from a cell line or cell strain that has previously been established.
  • Certain culture conditions depend on the cell type; however, each culture must consist of a suitable vessel with a substrate or medium that supplies the nutrients (such as amino acids, carbohydrates, vitamins, and minerals), growth factors or essential hormones for culturing cells. 
  • Gases (O2, CO2) and the physicochemical environment (pH, osmotic pressure, and temperature) also play an important role to regulate the proper cell growth in an artificial environment.

Working Principle of Cell Culture Virtual Lab

  • Cell culture is an amazing tool that allows for easy control and manipulation of all physicochemical and physiological cell factors, such as temperature, osmotic pressure, pH, gas, hormones, and media and its supplements. 
  • Media supplements help to optimize cell growth for specific applications depending on the chosen tissue or cell type. 
  • To be successful in cell culture, it is essential to remain in a contamination-free environment (bacteria, fungi, mycoplasma, etc.), so following aseptic techniques ensures that no microorganisms enter the cell culture.
  • In mammalian cell culture laboratory protocol, adherent cell lines will be first thawed from liquid nitrogen and seeded in a cell culture flask. 
  • Following 24 hours, cells will be visualized by light microscope for confluency, in order to define the best time for subculturing and scaling up of cells. 
  • Once the cells exceed 90% confluency, they will be subcultured and passed into other culture vessels. 
  • For long-term storage, cells will be frozen in a special freezing medium before storing them in liquid nitrogen.

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