Quantification of cell hybridoma yields with confocal microscopy and flow cytometry.
The fusion of antigen presenting and cancer cells leads to the formation of hybrid cells, which are considered a potential vaccine for treating cancer. The quality assessment of hybrid cell vaccines is crucial for the introduction of this new treatment. Flow cytometry was the method used recently, since it is faster in comparison to classical microscopy. Here we describe a rapid confocal microscopy based approach to quantify hybrid cell yields. The extent of fusion rate was determined by confocal microscopy by counting dual fluorescent cells and by measuring the area of co-localized pixels. Results of both methods showed high degree of correlation. The same samples were also analyzed by flow cytometry. Fusion rates determined with both techniques showed significant correlation. In conclusion, using confocal microscopy we developed a sensitive and a rapid method to assess the yield of hybridomas in a large number of electrofused cells.
Gabrijel
Mateja
M
Celica Biomedical Sciences Center, Stegne 21, 1000, Ljubljana, Slovenia.
Repnik
Urska
U
Kreft
Marko
M
Grilc
Sonja
S
Jeras
Matjaz
M
Zorec
Robert
R
D000818:Animals; D002459:Cell Fusion / Q000379:methods; D003713:Dendritic Cells / Q000166:cytology; D004558:Electric Stimulation; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000166:cytology* / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D016716:PC12 Cells; D051381:Rats; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013223:Statistics as Topic
D005456:Fluorescent Dyes