International Journal of Cell Cloning, Vol 1, 478-485, Copyright © 1983 by AlphaMed Press
A hypo-osmotic medium to disaggregate tumor cell clumps into viable and clonogenic single cells for the human tumor stem cell clonogenic assay
A Leibovitz, R Liu, C Hayes and SE Salman
A hypo-osmolar medium and tissue processing technique is described which is
useful for disaggregation of residual human tumor cell clumps persisting
after mechanical or enzymatic treatment of solid tumors and malignant
effusions. The addition of the hypo-osmolar procedure to the standard
methods for disaggregation increased the viable single cell yield in solid
tumors by 47% and in malignant effusions by 67%. In 5 of the 26 solid tumor
specimens tested in the human tumor stem cell assay, clonogenic single
cells were obtained with the hypo-osmolar procedure, whereas no growth was
observed using standard methods. Overall, the success rate for
clonogenicity increased from 46% to 65% for the 26 solid tumors, with the
major improvement occurring in ovarian cancer. Clonogenicity was obtained
in 80% of malignant effusions both by standard methods and the hypo-osmolar
techniques. The increased total yield of clonogenic cells obtained with
this procedure enhances the opportunity for experimental versatility and in
vitro drug testing.
