A novel method for synchronizing a B cell lymphoma.
Certain sub-lines of the murine B cell lymphoma BCL1 can be maintained in vitro and respond to cytokines including IL-2 and IL-5. BCL1 cells, as well as other B lymphomas, are difficult to synchronize using conventional techniques such as thymidine block or DNA synthesis inhibition. We have found that BCL1 cells maintained in Dulbecco's minimum essential medium (DMEM) with non-essential amino acids (NEAA) can be readily synchronized by culture in DMEM lacking NEAA. Within 10-18 h of medium replacement, 98% of BCL1 cells are 2 N in DNA content, suggesting that these cells are arrested in G0/G1. This population of BCL1 cells is viable and can be stimulated to enter S phase by culture in media containing NEAA; however, arrested cells did not appear to return synchronously into the cell cycle on addition of NEAA. A transient increase in levels of c-fos and c-myc mRNA was not detected after arrested BCL1 cells were stimulated to enter S phase, suggesting that arrested cells are in the G1 phase of the cell cycle, rather than G0. This technique for obtaining G1 arrested B lymphoma cells may prove useful in the analysis of molecular events that occur in B cells as a function of cell cycle position.
Isakson
P C
PC
Department of Pharmacology, University of Virginia Medical School, Charlottesville.
Purkerson
J
J
Catron
K
K
Bender
T P
TP
D000596:Amino Acids / Q000378:metabolism; D002453:Cell Cycle; D015870:Gene Expression; D016762:Genes, fos; D016259:Genes, myc; D006801:Humans; D066298:In Vitro Techniques; D007532:Isoleucine / Q000172:deficiency; D016393:Lymphoma, B-Cell / Q000473:pathology*; D016760:Proto-Oncogene Proteins c-fos / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D016760:Proto-Oncogene Proteins c-fos; D016271:Proto-Oncogene Proteins c-myc; D007532:Isoleucine