Combined flow and absorption DNA measurements of [3H]TdR-labelled tumour cells. I. Studies of MCa-11 cells grown as tumours in vivo and as exponential cultures in vitro.
Flow cytometry of cellular DNA content provides rapid estimates of DNA distributions, i.e. the proportions of cells in the different phases of the cell cycle. Measurements of DNA alone, however, yield no kinetic information and can make it difficult to resolve the cell cycle distributions of normal and transformed cells present in tumour biopsy specimens. The use of absorption cytophotometry of the Feulgen DNA content and [3H]TdR labelling of the same nuclei provides objective criteria to distinguish the ranges of DNA content for G0/G1, S, and G2/M cells. We now report on a study in which we combined flow and absorption cytometry to resolve the cell cycle distributions of host and tumour cells present in biopsy specimens of MCa-11 mouse mammary tumours labelled in vivo for 0.5 hr with [3H]TdR. A similar analysis of exponential monolayer cultures, labelled for 5 min with [3H]TdR under pulse-chase conditions, revealed a highly synchronous traversal of almost all cells through the different phases of the cell cycle. Combination of the flow and absorption methods also allowed us to detect G2 tumour cells in vivo and a minor tumour stem-line in vitro, to show that these two techniques are complementary and yield new information when they are combined.
Department of Surgery, Albuquerque Veterans Administration Medical Center, NM.
D C
DC
Allison
M
M
Chakerian
P F
PF
Ridolpho
S
S
Anderson
S
S
Curley
M E
ME
Wilder
J
J
Robertson
D000042:Absorption; D000818:Animals; D002453:Cell Cycle; D003592:Cytophotometry; D004273:DNA, Neoplasm / Q000032:analysis*; D005434:Flow Cytometry; D007399:Interphase; D007700:Kinetics; D008325:Mammary Neoplasms, Experimental / Q000032:analysis*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000032:analysis; D013936:Thymidine / Q000378:metabolism; D014316:Tritium; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D014316:Tritium; D013936:Thymidine