The selection of high-producing cell lines using flow cytometry and cell sorting.
The selection of high-producing cell lines is usually time-consuming and labour-intensive. Following transfection, high-producing cells are selected using limiting dilution cloning to prevent non- and low-producing cells from outgrowing high-producing cells, a process that normally takes > 3 months. During this time, the cells have to be screened occasionally to ensure stability of the selected clone. Several new methods for selecting and screening cells using flow cytometry and cell sorting have recently been developed; these include gel microdrop technology, which encapsulates the cells in gelatine beads, and matrix-based secretion assays. This paper reviews these techniques for selecting high-producing cell lines and isolating rare cells.
Carroll
Silvia
S
Department of Chemical Engineering, University of Birmingham, UK.
Al-Rubeai
Mohamed
M
D000818:Animals; D000937:Antigen-Antibody Reactions; D001360:Avidin; D019921:Biotinylation; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods*; D006224:Cricetinae; D004339:Drug Compounding; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis; D005780:Gelatin; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection
D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; C485192:green fluorescent protein, Aequorea victoria; D001360:Avidin; D049452:Green Fluorescent Proteins; D005780:Gelatin; D016650:Fluorescein-5-isothiocyanate