Methods for the estimation of the number and quality of animal cells immobilized in carbohydrate gels.
Rapid and reliable methods for the determination of survival, proliferation, and metabolic activity of immobilized cells in gels are described. The first method is based on an MTT assay that measures qualitatively and quantitatively the metabolic activity of the cells. The second method determines cell number by measuring the amount of DNA available for Feulgen staining. In the third method, two fluorescent dyes are used to differentially stain viable and dead cells. The fourth method involves the use of glutaraldehyde to protect the cells when melting the gel to facilitate hemocytometric count. The presented techniques should help to test the efficiency of the immobilization procedures and to monitor the growth and survival of immobilized cells.
al-Rubeai
M A
MA
Department of Microbiology, University of Surrey, Guildford, UK.
Musgrave
S C
SC
Lambe
C A
CA
Walker
A G
AG
Evans
N H
NH
Spier
R E
RE
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D002452:Cell Count / Q000379:methods*; D002455:Cell Division; D002470:Cell Survival; D004396:Coloring Agents / Q000378:metabolism; D004247:DNA / Q000032:analysis; D005404:Fixatives; D005456:Fluorescent Dyes; D005782:Gels; D005976:Glutaral; D006825:Hybridomas; D051379:Mice; D012394:Rosaniline Dyes; D012685:Sepharose; D013194:Staining and Labeling; D013778:Tetrazolium Salts; D013844:Thiazoles; D014343:Trypan Blue
D004396:Coloring Agents; C007722:Feulgen stain; D005404:Fixatives; D005456:Fluorescent Dyes; D005782:Gels; D012394:Rosaniline Dyes; D013778:Tetrazolium Salts; D013844:Thiazoles; D004247:DNA; D012685:Sepharose; C022616:thiazolyl blue; D014343:Trypan Blue; D005976:Glutaral