Therapy. Make proteins: commercial scale. Stem and cancer cells. Antibody production: monoclonals. Embryo culture. Primary Human and animal. Cell culture. Tissue Culture – Concepts. ➢ Primary culture: cells obtained from animal tissue. ➢ Passing: Process by which cells are often diluted and replaced in new vessels . PDF | Surachai Unchern and others published BASIC The development of animal cell culture can be traced back to the work of Ross.
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PDF | Background Cell culture technology has spread prolifically within a century, a variety of culture media has been designed. This review goes through the. that a low-level contamination in an animal cell culture can quickly lead to bacterial of cell types) and this marked the start of animal cell culture techniques. Animal Cell Culture CONCEPT AND APPLICATION Sheelendra M. Bhatt Alpha Science International Ltd. Oxford, U.K. CONTENTS Preface vii Abbreviations xvii .
Cells can be maintained in Petri dishes or flasks 25 cm2 or 75 cm2 which have the added advantage that the flasks can be gassed and then sealed so that a CO2 incubator need not be used. Tissue culture ware is always chosen to match the procedure.
However, glassware such as pipettes should be soaked in a suitable detergent, then passed through a stringent washing procedure with thorough soaking in distilled water prior to drying and sterilizing. Pipettes are often plugged with non-absorbent cotton wool before putting into containers for sterilizing.
Autoclave bags are available for loose items. Aluminum foil also makes good packaging material. Liquid Nitrogen Deep Freezer: Invariably for continuous and finite cell lines, samples of cultures will need to be frozen down for storage. They should be frozen in exponential phase of growth with a suitable preservative, usually dimethylsulfoxide DMSO.
Water Still or Reverse Osmosis Apparatus: A double distilled or reverse osmosis water supply is essential for preparation of media, and rinsing glassware. The pH of the double distilled water should be regularly checked as in some cases this can vary. Therefore, water from one source should be used. These are obtainable in various designs to allow a wide range of volumes to be filtered e.
The optimum conditions of factors like pH, osmotic pressure, etc. Phenol Red is added as a pH indicator of the medium.
There are two types of media used for culture of animal cells and tissues: a Natural Media: The natural media are the natural sources of nutrient sufficient for growth and proliferation of animal cells and tissues. Coagulant, such as plasma clots. It is now commercially available in the form of liquid plasma kept in silicon ampoules or lyophilized plasma. Plasma can also be prepared in the laboratory taking out blood from male fowl and adding heparin to prevent blood coagulation.
Biological fluids such as serum. It also contains the growth factors which promotes cell proliferation, cell attachment and adhesion factors. Tissue extracts for example Embryo extracts- Extracts from tissues such as embryo, liver, spleen, leukocytes, tumour, bone marrow etc. Different types of synthetic media can be prepared for a variety of cells and tissues to be cultured. Advantages of Serum in Culture Medium are: 1. Serum binds and neutralizes toxins, 2.
Serum contains a complete set of essential growth factors, hormones, attachment and spreading factors, binding and transport proteins, Contents 3. It contains the protease inhibitors, 4. It increases the buffering capacity, 5.
It provides trace elements. It is not chemically defined, and therefore, its composition varies a lot, b. It is sometimes source of contamination by viruses, mycoplasma, prions, etc. It increases the difficulties and cost of downstream processing, d.
It is the most expensive component of the culture medium. A pH below 6. The regulation of pH is done using a variety of buffering systems. All commercial media are formulated in such a way that their final osmolality is around mOsm.
Commonly the necessary amino acids include cysteine and tyrosine, but some non-essential amino acids may be needed. Glutamine is also required by most cell lines and it has been suggested that cultured cells use glutamine as an energy and carbon source in preference to glucose, although glucose is present in most defined media. A wide range of suitable preparations are available from relatively specific antibiotics, e. Types of Animal Cell Culture: i. Primary Culture: When cells are surgically removed from an organism and placed into a suitable culture environment, they will attach, divide and grow.
This is called a Primary Culture. There are two basic methods for doing this. This method is called Enzymatic Dissociation. Advantages of Primary Cell Culture: The major advantages of primary cultures are the retention of: 1.
This may result in the loss or damage of specific membrane receptors, damage to the integrity of the membrane, and loss of cellular products. Sub-Culturing: When the cells in the primary culture vessel have grown and filled up all of the available culture substrate, they must be Sub-cultured to give them room for continued growth. This is usually done by removing them as gently as possible from the substrate with enzymes. Some cell lines can be harvested by gently scraping the cells off the bottom of the culture vessel.
Cell Line Development. Pages Bioreactors for Mammalian Cells. Hydrodynamic Damage to Animal Cells. Monitoring of Cell Culture. Serum and Protein Free Media. Glycosylation in Cell Culture. Modelling of Mammalian Cell Cultures. Nishikant P. Shirsat, Niall J. Searching for Certainty in an Uncertain World.
Perfusion Processes. Stephan C. Laura A. Palomares, Mauricio Realpe, Octavio T. Metabolic Flux Analysis: Duarte, Manuel J.
Carrondo, Paula M. Alves, Ana P. Cell Immobilization for the Production of Viral Vaccines.