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Fig. 2. Differentiation potential of AGM cells. (A-C) Osteogenic cell
differentiation. After 10 days in osteogenic permissive medium, cell colonies
stained positive for ALP activity (A), and cultures expressed the osteoblastic
markers osteopontin and osteocalcin (B, lane 2). Neither marker was expressed
in freshly isolated AGM cells (B, lane1). Differentiated cultures also showed
an upregulation of the osteogenic marker Cbfa1 (B, lane 2). At day
21, mineralization of the cultures was observed with Alizarin Red (C). (D)
Adipogenic differentiation and colony formation. After stimulation with
adipogenic-inductive medium for one week, Pparg was upregulated (B,
lane 2) when compared with the expression of this marker in freshly isolated
AGM cells (B, lane 1). (E-H) Chondrogenic differentiation. After 21 days in
chondrogenic medium, cells formed several cartilaginous foci (E) with an
extracellular matrix rich in proteoglycans, as detected by Toluidine Blue
staining (F). Foci were also strongly positive for collagen type II, as shown
by the brown DAB staining (G). A secondary antibody control is shown in H.
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