Synchronization of 9L rat brain tumor cells by centrifugal elutriation

Cell Biophysics
P C KengK T Wheeler

Abstract

Asynchronous 9L cells were separated into relatively homogeneously-sized populations using centrifugal elutriation with both a conventional collection method and a long collection method. A substantial increase in the homogeneity of the volume distributions and in the degree of synchrony of the separated fractions was obtained using the long collection method. Autoradiographic data indicated that fractions containing greater than or equal to 97% G1 cells, greater than or equal to 80% S cells, and 70-75% G2 cells could be routinely recovered with this procedure. Recovery in these fractions varied from 5 to 8% of the total number of cells elutriated. The colony forming efficiency (CFE) of cells from fractions representing each phase of the cell cycle was a constant 60-70%, which was comparable to the 60-80% usually found for asynchronous 9L cells. The percentage of cells in the G1, S, and G2 phases in the elutriated fractions was more accurately determined from the volume distribution than from computer fits of the DNA histogram obtained from flow cytometry. In genereal, the degree of synchrony was related to the coefficient of variation (CV) of the volume distributions of the elutriated fractions. The CV was about 14% for all el...Continue Reading

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Citations

Jan 1, 1992·Radiation and Environmental Biophysics·K T WheelerC A Wallen
Jul 1, 1987·International Journal of Radiation Biology and Related Studies in Physics, Chemistry, and Medicine·J P FreyerM R Raju
Jun 1, 1988·International Journal of Radiation Oncology, Biology, Physics·C M WestR M Sutherland
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Jan 1, 1984·Critical Reviews in Clinical Laboratory Sciences·J D Defreese, T S Wang
Jul 10, 2020·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Marie GoeppMarie-Paule Vasson

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