Biology of the human malignant lymphomas I. Establishment in continuous cell culture and heterotransplantation of diffuse histiocytic lymphomas

Abstract

With the aid of a new methodology of tissue culture which permits rapid screening of nutrient requirements, the tumor cells from pleural or peritoneal effusions of three patients with diffuse histiocytic lymphomas have been established in continuous culture in vitro and designated as the SU-DHL-I, SU-DHL-2, and SU-DHL-3 cell lines, respectively. All three cell lines required a 10% supplement of human pleural effusion fluid or human serum for continuous growth. Morphologically, they closely resembled the tumor cells of the original patients as seen in lymph node biopsies or in pleural or peritoneal effusion fluids. Histochemical studies revealed positive reactions for nonspecific esterase, acid phosphatase, and methyl green-pyronin in both the original tumor cells and the histiocytic lymphoma cell lines. Cytoplasmic vacuoles, which were particularly conspicuous in the-SU-DHL-I cells, were shown to contain lipid. In contrast, a lymphoblastoid cell line, SU-LB-I, was negative for nonspecific esterase, acid phosphatase, and lipid. The histiocytic lymphoma cell lines were more rigorously distinguishable from lymphoblastoid cell lines by their negative EBNA test reactions for the presence of the Epstein-Barr virus (EBV) genome and by their lack of surface immunoglobulin staining reactions characteristic of B-lymphocytes. Cytogenetic studies demonstrated that early culture passages of the histiocytic lymphoma cell lines were aneuploid. These cells were also successfully heterotransplanted in the brains of congenitally athymic (“nude”) mice, providing further evidence of their neoplastic character. These histiocytic lymphoma cell lines provide an important new resource for studies of this group of highly malignant human lymphomas.