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Biochemical and functional characterization of proteoglycans produced by SI/SI d murine bone marrow stromal cell lines
Author(s) -
Bentley Stuart A.,
Kirby Suzanne L.,
Anklesaria Pervin,
Greenberger Joel S.
Publication year - 1990
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041450109
Subject(s) - stromal cell , cell culture , bone marrow , extracellular matrix , proteoglycan , haematopoiesis , chemistry , glycosaminoglycan , microbiology and biotechnology , biology , biochemistry , immunology , stem cell , cancer research , genetics
The Steel anemia of mice results from an inherited defect in the hematopoietic microenvironment. Proteoglycans synthesized by bone marrow stromal cells are an important functional component of the hematopoietic microenvironment in normal animals. It is thus possible that Steel anemia results from a molecular abnormality involving bone marrow stromal proteoglycans. To investigate this possibility, we studied proteoglycan synthesis in three stromal cell lines from Steel anemic (SI/SI d ) animals and two control stromal cell lines, one (+/+2.4) from a non‐anemic littermate, and one (GBI/6) from a normal mouse. Proteoglycans were precursor labelled with 35 S sulfate and separated by ion exchange HPLC, CsCI density gradient centrifugation, and molecular sieve HPLC. Glycosaminoglycan (GAG) moieties were characterized by molecular sieve HPLC and enzyme sensitivity. There were no consistent differences in total proteoglycan synthesis, proteoglycan heterogeneity, GAG hydrodynamic size, or enzyme sensitivity among the cell lines studied. Growth factor binding to stromal extracellular matrix (ECM) was studied by co‐culture of an IL‐3‐dependent cell line (FDC‐P1) with cell‐free ECM preparations from an SI/SI d and a control (GBI/6) stromal cell line, with and without pre‐incubation with IL‐3. Cell‐free ECM preparations from SI/SI d and control cell lines supported FDC‐P1 growth to an approximately equal extent after pre‐incubation with II‐3. FDC‐P1 growth support by ECM preparations from both cell lines was also observed without IL‐3 pre‐incubation, although to a lesser extent, suggesting ECM binding of endogenous growth factors synthesized by the stromal cells.