Osteochondrogenic potential of marrow mesenchymal progenitor cells exposed to TGF-beta 1 or PDGF-BB as assayed in vivo and in vitro.
Mesenchymal progenitors cells can be isolated from rat bone marrow and mitotically expanded in vitro. When these cells, which we operationally call mesenchymal stem cells (MSCs), are placed in an appropriate environment, they have the capacity to differentiate into bone and/or cartilage. This capacity is called osteochondrogenic potential. In this study, preconfluent MSCs were exposed in vitro to 5 ng/ml transforming growth factor-beta 1 (TGF-beta 1) or platelet-derived growth factor, isoform BB (PDGF-BB) for a pulse of 48 h and assayed for cell proliferation, alkaline phosphatase activity, and osteochondrogenic potential; untreated MSC's served as controls. In these cell culture conditions, TGF-beta 1 or PDGF-BB had similar effects on proliferation and alkaline phosphatase activity. Both growth factors increased cell proliferation and decreased alkaline phosphatase activity of MSCs. Sister cultures of TGF-beta 1- or PDGF-BB-treated MSCs and untreated MSCs were trypsinized. For each type of culture, the trypsinised MSCs were split in two parts: one part was replated in an osteogenic medium to assess its in vitro osteogenic potential, whereas the other part was seeded into porous calcium phosphate ceramics and implanted subcutaneously in syngeneic rats to assess its in vivo osteochondrogenic potential. PDGF-pretreated MSCs showed no difference in in vivo and in vitro osteochondrogenesis from that of control MSCs, while TGF-beta 1 pretreatment blocked the osteochondrogenic potential of MSCs when assayed in vitro for bone nodule formation. However, when tested in vivo, TGF-beta 1-pretreated MSCs were able to form bone and cartilage. These data show that measurements of proliferation and alkaline phosphatase activity of preconfluent MSCs immediately after exposure to growth factor were not predictive of their subsequent osteochondrogenic potential. Moreover, the variation of the osteochondrogenic potential of MSCs after exposure to growth factor was further modulated by the environment in which the MSCs were assayed.
Cassiede
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Department of Biology, Case Western Reserve University, Cleveland, Ohio, USA.
Dennis
J E
JE
Ma
F
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Caplan
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AI
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000077214:Becaplermin; D001846:Bone Development / Q000187:drug effects; D001853:Bone Marrow / Q000187:drug effects* / Q000201:enzymology; D001854:Bone Marrow Cells; D002130:Calcium Phosphates / Q000737:chemistry; D002356:Cartilage / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002469:Cell Separation; D017690:Cell Transplantation; D002516:Ceramics; D006801:Humans; D008297:Male; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D016062:Porosity; D020574:Proto-Oncogene Proteins c-sis; D051381:Rats; D011916:Rats, Inbred F344; D011994:Recombinant Proteins / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism
D002130:Calcium Phosphates; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; D000077214:Becaplermin; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; D000469:Alkaline Phosphatase; D014357:Trypsin; C485829:calcium phosphate, dibasic, anhydrous