Development of a lacZ marked WEHI-3B D+ murine leukemic cell line as an in-vivo model of acute non-lymphocytic leukemia

Clay Smith, Marcus O. Muench, Marianne Knizewski, Eli Gilboa, Malcolm A.S. Moore

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Established leukemic cell lines have been useful models for studying the biology of leukemia. Analysis of the actions of differentiating agents on leukemic cell lines in vivo has been limited by an inability to unambiguously distinguish host hematopoietic elements from differentiated leukemic cells. In order to identify and quantify leukemic cells during in vivo studies, a derivative of the murine myelomonocytic leukemia cell line WEHI-3B D+, which stably expresses β-galactosidase, was constructed utilizing retroviral vector gene transfer. This cell line, termed WEHI-3B D+/lacZ 2.8, demonstrated in vitro growth and differentiation properties similar to the parental cell line. WEHI-3B D+/lacZ 2.8 expressed high levels of β-galactosidase following prolonged in vitro growth and following differentiation in suspension cultures and clonogenic assays. In vivo, WEHI-3B D+/lacZ 2.8 was leukemogenic and high level expression of β-galactosidase was maintained. Quantification of tissue involvement with WEHI-3B D+/lacZ 2.8 leukemia was performed utilizing staining with the fluorogenic β-galactosidase substrate fluorescein di-β-galactoside and fluorescence-activated cell sorting analysis. In vivo differentiation efficiency following granulocyte colony-stimulating factor (G-CSF) administration was determined using a simultaneous nuclear and cytoplasmic staining procedure. Results indicate that treatment of mice inoculated with WEHI-3B D+/lacZ 2.8 cells with G-CSF administration causes detectable but limited differentiation.

Original languageEnglish (US)
Pages (from-to)310-317
Number of pages8
JournalLeukemia
Volume7
Issue number2
StatePublished - Feb 1993
Externally publishedYes

ASJC Scopus subject areas

  • Hematology
  • Oncology
  • Cancer Research

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