Oligonucleotide fingerprinting as a means to identify and survey long-term cultured B cell hybridomas and T cell lines.
Common problems encountered during cell culture are cross-contamination, instability, and inadvertent exchange of cells. Here we report on the application of oligonucleotide fingerprinting as a simple and efficient method to screen hybridomas and T cell lines. Among the fingerprint probes tested, the simple repetitive oligonucleotide (CAC)5/(GTG)5 proved to be most useful for obtaining many fragments specific for each cell line. Because of variable loss of chromosomes, cloned hybridoma cells from one fusion exhibit different fingerprint patterns. Thus, antibody-secreting B cell hybridomas can be distinguished easily even when they originate from the same fusion. Furthermore, we were able to monitor the genomic integrity of myelomas and hybridomas over a period of more than 2 years, thereby highlighting long-term stability. Another application of the method is the control of T cell lines requiring irradiated or mitomycin-treated feeder cells for continuous growth or cloning. There cell lines are always threatened to be overgrown by feeder cells that have escaped from the lethal pretreatment. T cell clones of one individual, known to display differently rearranged T cell receptors, did not show differences in their fingerprint patterns. However, in EBV-transformed cloned B cells, slight differences between clones from the same donor were identified.
Hampe
J
J
Institute for Medical Immunology, Faculty of Medicine (Charité), Humboldt University, Berlin, Germany.
Nürnberg
P
P
Epplen
C
C
Jahn
S
S
Grunow
R
R
Epplen
J T
JT
D001402:B-Lymphocytes / Q000166:cytology*; D001483:Base Sequence; D002460:Cell Line; D002472:Cell Transformation, Viral; D004247:DNA / Q000235:genetics; D016172:DNA Fingerprinting / Q000379:methods*; D004854:Herpesvirus 4, Human; D006801:Humans; D006825:Hybridomas / Q000166:cytology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D013601:T-Lymphocytes / Q000166:cytology*
D015345:Oligonucleotide Probes; D004247:DNA