Detection of surface differences between closely related cell populations by partitioning. Cultured K-562 cell sublines.
The K-562 cell line is a culture of human leukemia stem cells originally derived from a patient with chronic myelogenous leukemia in blast crisis. We have applied a sensitive method capable of detecting subtle differences in charge-associated and noncharge-related cell surface properties between closely related cell populations to K-562 cells from different sources and having different histories. The method consists of isotopically labeling aliquots of each of two cell populations to be compared with 51Cr-chromate and mixing the labeled cells with an excess of unlabeled cells with which they are to be compared. The mixtures are subjected to countercurrent distribution in either a charge-sensitive or a noncharge-sensitive dextran-poly(ethylene glycol) aqueous two-phase system. The distribution curves are analyzed for total cells (in terms of electronic counts) and labeled cells (in terms of cpm). Alterations in relative specific activities through the distribution curves are indicative of differences in surface properties between such cell populations. Using this method we have found surface differences, both charge-associated and noncharge-related, between any two K-562 cell sublines examined. Interestingly, whereas we observed differences among K-562 sublines, we never witnessed a change in surface properties of the respective sublines. The differences among the sublines examined remained unaltered for more than 40 passages in our hands. It thus appears likely that the event(s) leading to an altered K-562 cell surface, detectable by partitioning, does not occur gradually.
Walter
H
H
Laboratory of Chemical Biology, Veterans Administration Medical Center, Long Beach 90822.
Krob
E J
EJ
Al-Romaihi
F A
FA
Johnson
D
D
Lozzio
C B
CB
D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure*; D002469:Cell Separation / Q000379:methods*; D003377:Countercurrent Distribution; D006801:Humans; D007553:Isotope Labeling; D007942:Leukemia, Experimental / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000648:ultrastructure*