By Shirley McBride, Mary Heenan, Martin Clynes (auth.), Dr. Martin Clynes (eds.)
Cell tradition ideas let a number of molecular and phone organic inquiries to be addressed, providing physiological stipulations while warding off using laboratory animals. as well as uncomplicated suggestions, a variety of specialized sensible protocols overlaying the next parts are incorporated: cellphone proliferation and demise, in-vitro versions for cellphone differentiation, in-vitro types for toxicology and pharmacology, business software of animal cellphone tradition, genetic manipulation and research of human and animal cells in culture.
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Extra info for Animal Cell Culture Techniques
Allow the broth mixture to cool to room temperature. 4. Aseptically add 50 ml serum, 25 ml yeast extract (15 % w/v) and 25 ml stock solution. Mix the solution completely. 4. 5. Dispense 5 ml aliquots of the broth medium into sterile test tubes and store at 4°C, Use within 3 -4 weeks. Mycoplasma Agar Medium 1. 95 g mycoplasma agar base to 150 ml water. To dissolve, bring the solution to the boil. 2. Sterilize the solution by autoclaving. 3. Place the sterilized medium in a water bath at 50°c' Place 50 ml serum, 25 ml yeast extract (15 % w/v) and 25 ml stock solution in a water bath at 37°C.
Slow growing cells will have a low split ratio. 2. 1. 2. Split ratio Cell type Split ratio African green monkey kidney (Vero) Fetal rhesus kidney (FRK) Human epithelium (Hep-2) Human embryonic lung (MRC-5) Human rhabdomyosarcoma (RD) Madin Darby canine kidney (MDCK) 1:3 1:2 1:5 1:2 1:2 1:4 2 Cell Culture and Diagnostic Virology 21 Anchorage-dependent cells are dispersed from the monolayers using equal volumes (10 ml) of a chelating agent, versene (EDTA), and a proteolytic enzyme, trypsin. Cells may also be removed by scraping with a disposable cell scraper or by using sterile glass beads.
Catalogue number 30-500-44). These treatments have been used to various effects by different researchers in the eradication of mycoplasma from cell stocks (Fleckenstein et al. 1994; Somasundaram et al. 1992; Hlubinova et al. 1993; Nissen 1995; Drexler et al. 1994). 52 MARY HEENAN, MARTIN CLYNES Antibiotic treatment of mycoplasma contamination should last until the contamination is completely removed. This should be determined by negative results in mycoplasma tests, after the cultures have been grown in treatment-free medium for at least 4 weeks.
Animal Cell Culture Techniques by Shirley McBride, Mary Heenan, Martin Clynes (auth.), Dr. Martin Clynes (eds.)