Use of deoxyribonucleic acid probes in the identification of cell origin and detection of cellular contamination in human lymphoblastoid cell lines.
Established lymphoblastoid cell lines have provided a valuable reference source for studying neoplastic lymphoproliferative disorders in humans. However, two major problems are associated with the establishment and growth of these cell lines: (a) the established cell line may not represent the original neoplastic clone, and (b) contamination of the established cell line with the other cell lines may occur. Lymphoblastoid cell lines "W" and "SP5" were established from splenectomy specimens of two patients with hairy cell leukemia. Both cell lines displayed B cell characteristics by immunophenotypic and Ig gene rearrangement studies. The banding patterns of the rearranged Ig genes (heavy and light chains) in the W cell line were different and in the SP5 cell line were identical with the corresponding untransformed splenic cell lines, indicating that cell line SP5 did and cell line W did not represent the original neoplastic clone. Continuous cultures of some of the subclones derived from cell line W and SP5 led to the growth of the cell lines W15T, W17T, and SP5T which all demonstrated T cell features based on immunophenotypic and T cell receptor rearrangement studies. However, the T cell receptor alpha and beta rearranged bands as well as bands generated by hybridization with highly polymorphic DNA probes p YNH24 and 0-3315-32 in these three lines and a human T cell leukemia line (CEM), were identical indicating that W15T, W17T and SP5T cell lines were contaminated with CEM. Studies of gene establishment patterns and DNA polymorphisms by Southern blotting are effective methods to establish clonal identity and to rule out cellular contamination in lymphoblastoid cell lines.
Naeim
F
F
Department of Pathology, University of California, Los Angeles.
Jensen
L
L
Susi
E
E
Yam
P
P
Gatti
R
R
Salser
W
W
Concannon
P
P
D001402:B-Lymphocytes; D015139:Blotting, Southern; D015342:DNA Probes; D015321:Gene Rearrangement; D015329:Gene Rearrangement, T-Lymphocyte; D005803:Genes, Immunoglobulin; D006680:HLA Antigens / Q000145:classification; D006801:Humans; D007943:Leukemia, Hairy Cell / Q000473:pathology*; D008213:Lymphocyte Activation; D011110:Polymorphism, Genetic; D013601:T-Lymphocytes; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D006680:HLA Antigens