Recent isolates of RX54-3 hybridoma cells (new cells) protect BALB/c mice against subsequent challenge with the tumorigenic myeloma parent cells used to construct this hybridoma. In contrast, hybridoma cells which have been maintained in tissue culture for long periods of time (old cells) are not protective. In the present study, we compared a number of properties of the new and old hybridoma cells and determined which line was more similar to the parent myeloma. We found that new hybridoma cells resembled myeloma cells in: (a) possessing A- and C-type viral particles on transmission electron microscopy and a relatively smooth surface on scanning electron microscopy; (b) being sensitive to a hypotonic solution containing the dye propidium iodide; (c) having similar DNA histograms on flow cytometric analysis; (d) being sensitive to the bacteriocin colicin HSC 10; and (e) being tumorigenic in nude mice. In contrast, old hybridoma cells differed in all of these characteristics from new hybridoma and myeloma cells. Therefore, in order to protect against challenge with the tumorigenic myeloma parent, hybridoma cells must retain properties of that parent.
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