Characterization of Murine Stromal Cell Clones Established From Bone Marrow and Spleen

Leukemia. 1996 May;10(5):803-12.

Abstract

Stromal cell lines were established by irradiating adherent layers of bone marrow and spleen cells in Dexter-type long-term culture with X-rays. Some of these cell lines support myelopoiesis and/or B lymphopoiesis in vitro. Furthermore, the characteristics of these stromal cell lines were studied. Cytokine activity was detected in the conditioned media from all hematopoietic-supportive and non-supportive stromal cells. Quantitative reverse transcriptase-polymerase chain reaction analysis revealed that the mRNAs of macrophage colony-stimulating factor and stem cell factor, but not that of Interleukin-3, were detectable in all the hematopoietic-supportive and non-supportive stromal cell lines. The transcripts of granulocyte colony-stimulating factor, interleukin-6, interleukin-7, and leukemia inhibitory factor were expressed in a wide variety of cell lines. Most stromal cell lines synthesized mRNA of c-mpl, the ligand of which stimulates megakaryopoiesis and thrombopoiesis. These observations indicate that the pattern of mRNA expression of cytokines is not correlated with the hematopoietic-supportive ability of stromal cell lines. There was a significant difference in the efficiency of adhesion of lineage marker-negative bone marrow cells to fibroblasts and stromal cell lines. This appears to be correlated with the hematopoietic-supportive ability of the stromal cell lines.

MeSH terms

  • 3T3 Cells
  • Animals
  • B-Lymphocytes / cytology
  • Base Sequence
  • Biomarkers
  • Bone Marrow Cells*
  • Cell Adhesion
  • Cell Line
  • Cell Lineage
  • Coculture Techniques
  • Connective Tissue / physiology
  • Connective Tissue Cells*
  • Fibroblasts / metabolism
  • Gene Expression
  • Hematopoiesis / physiology
  • Hematopoietic Cell Growth Factors / biosynthesis
  • Hematopoietic Cell Growth Factors / genetics
  • Hematopoietic Stem Cells / radiation effects
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C3H
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Spleen / cytology*

Substances

  • Biomarkers
  • Hematopoietic Cell Growth Factors
  • RNA, Messenger