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Unformatted text preview: dy temperature of the host from which
the cells were obtained. With cold-blooded vertebrates, a temperature range of 18° to 25°C
is suitable; most mammalian cells require 36° to 37°C. This temperature range is usually
maintained by use of carefully calibrated, and frequently checked, incubators. Basic environmental
Q Q Q Controlled
Good substrate for
and incubator that
maintains the correct
pH and osmolality Anchorage-dependent cells also require a good substrate for attachment and growth. Glass
and specially treated plastics (to make the normally hydrophobic plastic surface hydrophilic
or wettable) are the most commonly used substrates. However, Attachment Factors, such
as collagen, gelatin, fibronectin and laminin, can be used as substrate coatings to improve
growth and function of normal cells derived from brain, blood vessels, kidney, liver, skin,
etc. Often normal anchorage-dependent cells will also function better if they are grown on a
permeable or porous surface. This allows them to polarize (have a top and bottom through
which things can enter and leave the cell) as they do in the body. Transwell® inserts are
Corning vessels with membrane-based permeable supports that allow these cells to develop
polarity and acquire the ability to exhibit special functions such as transport. Many specialized cells can only be truly “happy” (function normally) when grown on a porous substrate
in serum-free medium with the appropriate mixture of growth and attachment factors.
Cells can also be grown in suspension on beads made from glass, plastic, polyacrylamide and
cross-linked dextran molecules. This technique has been used to enable anchorage-dependent
cells to be grown in suspension culture systems and is increasingly important for the manufacture of cell-based biologicals. Corning® Transwell permeable
supports are used to study cell
transport and migration. The culture medium is the most important and complex factor to control in making cells
“happy”. Besides meeting the basic nutritional requirement of the cells, the culture medium
should also have any necessary growth factors, regulate the pH and osmolality, and provide
essential gases (O2 and CO2). The ‘food’ portion of the culture medium consists of amino
acids, vitamins, minerals, and carbohydrates. These allow the cells to build new proteins and
other components essential for growth and function as well as providing the energy necessary for metabolism. For additional information on this topic, see the article: Construction
of Tissue Culture Media by C. Waymouth in Growth, Nutrition and Metabolism of Cells in
Culture, Volume 1, (1972; Ref. 5).
The growth factors and hormones help regulate and control the cells’ growth rate and functional characteristics. Instead of being added directly to the medium, they are often added in
an undefined manner by adding 5 to 20% of various animal sera to the medi...
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This note was uploaded on 09/30/2013 for the course PHARM 101 taught by Professor Mishra during the Fall '11 term at Birla Institute of Technology & Science, Pilani - Hyderabad.
- Fall '11