A novel method for synchronizing a B cell lymphoma

J Immunol Methods. 1991 Dec 15;145(1-2):137-42. doi: 10.1016/0022-1759(91)90319-b.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acids / metabolism
  • Cell Cycle
  • Gene Expression
  • Genes, fos
  • Genes, myc
  • Humans
  • In Vitro Techniques
  • Isoleucine / deficiency
  • Lymphoma, B-Cell / pathology*
  • Proto-Oncogene Proteins c-fos / genetics
  • Proto-Oncogene Proteins c-myc / genetics
  • Tumor Cells, Cultured

Substances

  • Amino Acids
  • Proto-Oncogene Proteins c-fos
  • Proto-Oncogene Proteins c-myc
  • Isoleucine