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Preliminary characterization of porcine bone marrow stromal cells: Skeletogenic potential, colony‐forming activity, and response to dexamethasone, transforming growth factor β, and basic fibroblast growth factor
Author(s) -
Thomson B.M.,
Bennett J.,
Dean V.,
Triffitt J.,
Meikle M.C.,
Loveridge N.
Publication year - 1993
Publication title -
journal of bone and mineral research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.882
H-Index - 241
eISSN - 1523-4681
pISSN - 0884-0431
DOI - 10.1002/jbmr.5650081004
Subject(s) - stromal cell , transforming growth factor , bone marrow , growth factor , fibroblast growth factor , endocrinology , human bone , biology , medicine , microbiology and biotechnology , cancer research , immunology , in vitro , genetics , receptor
Neonatal pig bone marrow stromal cells (PBMSC) were tested in vivo and in vitro to establish their use as a large‐animal model for the study of skeletogenesis. When implanted in diffusion chambers in athymic mice for 6–8 weeks, both freshly isolated pig bone marrow and passage 2 PBMSC formed partially mineralized cartilage, bone‐like material, and fibrous tissue. The cartilage showed metachromatic, perilacunar staining with toluidine blue and safronin O, alcian blue staining for chondroitin and keratan sulfate, and intense immunostaining for type II collagen. Osteocalcin was immunolocalized to the mineralized regions, consistent with the formation of bone. Alkaline phosphatase was primarily observed in cell layers at boundaries between tissue types. Unstimulated monolayer cultures of PBMSC produced type I but not type II collagen, responded to dexamethasone (10 −8 M) with a 1.7‐fold increase in alkaline phosphatase activity, and were stimulated to divide by basic fibroblast growth factor (1.5‐fold; EC 50 1 ng/ml). Transforming growth factor β (TGF‐β) blocked both dexamethasone‐induced alkaline phosphatase expression (EC 50 , 1 ng/ml of TGF‐β) and the mitogenic effects of bFGF (EC 50 0.06 ng/ml of TGF‐β). When incubated for 10–14 days in medium containing dexamethasone, β‐glycerophosphate and ascorbate PBMSC formed mineralized nodules. Calcification occurred in the middle of the aggregates and was associated with intensely alkaline phosphatase positive cells and a dense type I collagen‐rich matrix. PBMSC also displayed colony‐forming unit‐fibroblastic activity, with approximately 1 in 80 of the plated cells formed colonies > 128 cells over 14–21 days. PBMSC therefore mimic the known activities of stromal cells from other species, including the human, suggesting that they are a valid model for skeletal research.