Distinct hematopoietic support by two human stromal cell lines

Exp Hematol. 2001 Jun;29(6):736-45. doi: 10.1016/s0301-472x(01)00644-0.

Abstract

Objective: The hematopoietic microenvironment is complex, and the role of myofibroblast in its function is crucial. In order to obtain a stable model reflecting this particular cell type, we have previously established human bone marrow cell lines from primary myofibroblastic Stro1(+) population (pStro1(+)). We placed HPV16 E6 and E7 expression under the control of different promoters. Here, we have characterized and studied the hematopoietic support for two cell lines corresponding to the promoters alpha-SM (alphaSM-56 line) and SV40 (SV40-56 line).

Materials and methods: The expression profile was analyzed at the RNA level by gene array and at the protein level by Western blot, flow cytometry, and ELISA. Hematopoietic support determined using colony-forming unit (CFU) and stroma-adherent colony-forming cell (SA-CFC) assays.

Results: The phenotype of cell lines was not significantly modified compared with primary myofibroblastic cells. They secreted a broad spectrum of hematopoietic cytokines and nonspecific mediators. The two lines allowed the growth of hematopoietic precursors and had different support capabilities.

Conclusions: We have extensively characterized two novel human bone marrow stromal cell lines. They retained a myofibroblastic phenotype and have substantial but different hematopoietic support capabilities. These lines provided a basis for determining stromal factors involved in stem-cell regulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / analysis
  • Bone Marrow Cells / cytology
  • Cell Adhesion
  • Cell Adhesion Molecules / analysis
  • Cell Line
  • Cells, Cultured
  • Coculture Techniques
  • Colony-Forming Units Assay
  • Cryopreservation
  • Culture Media
  • Cytokines / analysis
  • Cytokines / genetics*
  • Flow Cytometry
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / physiology*
  • Humans
  • Integrins / analysis
  • Phenotype
  • Stromal Cells / cytology
  • Stromal Cells / physiology*

Substances

  • Antigens, CD34
  • Cell Adhesion Molecules
  • Culture Media
  • Cytokines
  • Integrins