CELL CYCLE SYNCHRONIZATION OF MOUSE LIVER EPITHELIAL CELLS BY ELUTRIATION CENTRIFUGATION
Detailed methods are described for the sorting and cell cycle synchronization by means of centrifugal elutriation of an established mouse liver epithelial cell line(NMuLi). In a comparison between three different elutriation media and between two different temperatures(4° and 20° C), the NMuLi cells were found to be most reproducibly sorted in the cell cycle when run in growth medium in the absence of serum and at the lower temperature. Under these conditions. and using decrements of rotor speed calculated from an empirically derived algorithm as described in the text an initially asynchronous population (38% G{sub 1}, 36% S, and 28% G{sub 2}M) was sorted into fractions enriched to 60% G{sub 1}, 75% S, and 50% G{sub 2}M. Of the cells loaded into the rotor, 30% were lost in the elutriation process, and about 20% recovered as aggregates. The remainder appeared in the various synchronized fractions. Epithelial cells sorted in this manner demonstrated no loss of viability, and upon replating showed significant movement in the cell cycle by 6 hrs post elutriation. The degree of synchronous movement through the cell cycle achieved by elutriation depended on the part of the cell cycle from which the original elutriated fraction came. Cells collected as late S and G{sub 2}M moved through the cell cycle with the tightest sychrony.
- Research Organization:
- Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Sponsoring Organization:
- Chemical Sciences Division
- DOE Contract Number:
- DE-AC02-05CH11231
- OSTI ID:
- 1072212
- Report Number(s):
- LBL-11067
- Journal Information:
- Cell and Tissue Kinetics, Journal Name: Cell and Tissue Kinetics
- Country of Publication:
- United States
- Language:
- English
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