Adaptation of human long-term B lymphoblastoid cell lines to chemically defined, serum-free media

Abstract

Attempts were made to adapt human long-term B lymphoblastoid cell lines to prolonged growth in serum-free, chemically defined media. A newly described medium, which is an enriched modification of Dulbecco's modified Eagle's medium containing additional amino acids and vitamins, was used. The serum is totally replaced by albumin, transferrin, and soybean lipid. The cell lines were all adaptable from RPMI 1640 over a period of time during which the 10% fetal bovine serum (FBS) concentration was reduced and then eliminated in successive steps. After 3 to 6 wk minor alterations in cell shape and adhesion were noted without significant histological changes. Growth characteristics were comparable in the new medium provided a double initial inoculum was used. A panel of cell surface markers, including surface immunoglobulins, Ia antigens, Fc and complement receptors, and T and B erythrocyte rosettes, all showed no altered expression. Molecular genotyping of Ia antigens was carried out by 3-D gel electrophoresis. The antigens showed their full polymorphism without change and were shed into the new culture medium without alteration. Chromosome analysis was performed on Q-banded karyotypes from one of the lines and showed no alteration resulting from the change to serum-free conditions. Thus long-term B lymphoblastoid cell lines can be adapted to prolonged growth in serum-free medium. This will facilitate the assay and isolation of cell products regulating lymphocyte function and the identification and characterization of cell surface molecules free of interference from undefined serum components.

Characterization of A U‐937 subline which can be induced to differentiate in serum‐free medium

“…The overall expression of surface antigens, including the TfR, was not altered by adaptation to serum-free conditions. This is in agreement with the observations of Muzik et al (1982) who found no alterations in surface antigen expression of B-lymphoblastoid cell lines adapted to growth in a serumfree, defined medium. However, the significance of the reduced expression of lactosyl ceramide (CDwl7), complement receptor 1 (CD35) and CD38 observed in the U-937-1SF cells is at present unclear.…”