Chapter 1 – Introduction
  Textbook: „Introduction to Mammalian Cell Culture“       
  (D. Ler, U. Glamočlija, M. Suljagić)
   Introduction
   • Cell Culture – removal of cells from an organism and their 
    subsequent growth in a favorable, artificially controlled 
    environment
   • Primary Cell Culture – removal from tissue
   • Can be sub-cultured
   • Cell line – due to proliferation
    • Finite – limited life span
    • Immortal – due to transformation (Continous cell line)
   Primary Cell Culture
   • Contains heterogenous cell population
   • Sub-culturing leads to cell line generation
   • Different purification methods
   • Tissue sample: Dissection, Rinsing, Mechanical 
    Dissagregation (Enz. Digestion) in trypsin
   • Different separation techniques:
    • Density gradient separation
    • MACS
    • FACS
                            Figure – Cell line
   • Sub-culturing – cells are harvested, diluted in a fresh growth 
   medium and replaced in a new culture flask to promote further 
   growth
   • Essential for viable state
   • PRIMARY CUTURE  Sub-culturing  CELL LINE 
   (SEC.CULTURE)
   • Cell line – transfer of cell from one to another culture vessel
   • Providing fresh nutrients and growing space
         Figure – Primary Cell Culture
      Types of cell lines (on the basis of life span of culture):
      1.  Finite cell lines - limited life span
                           - limited number of cell generations (40-60 doublings)
                           - slow growth rate
                           - doubling time 24-96 h
      2. Continuous cell lines  - transformed under laboratory conditions
                                                - grow in a monolayer or in suspension
                       - rapid growth rate
                       - doubling time 12-24 h 
      • Cell strain – positive selection of cells either from primary culture or 
       cell line by cell cloning
      • Limited division potential, lose ability to proliferate (Hayflick limit)