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. 2002 Jun 17;195(12):1549–1563. doi: 10.1084/jem.20011700

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Copyright © 2002, The Rockefeller University Press

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Figure 4. — Osteoblasts, adipocytes, and chondrocyte differentiation. (A) Total LMCs (a, d, and g), ALCAM^low/− cells (b, e, and h), and ALCAM^high cells (c, f, and i) were induced (d–i) or not-induced (a–c) to undergo osteogenic differentiation. After 2 wk, ALP staining was performed. Panels g–i show high power fields of d–f, respectively. Scale bars, 1 mm (a–f), 200 μm (g–i). (B) The region of alizarin red positive calcium deposition in ALCAM^high cells, in which osteogenic differentiation was induced for 2 wk. (a) no factor; (b and c) β-GP and ascorbic acid. (b) The center of culture plate. (c) The corner of the culture plate. (C) TRAP activities were measured on day 7 of cocultivation of BM derived osteoclast precursor cells and ALCAM^high cells, which were induced (hatched bar) or not induced (solid bar) to undergo osteogenic differentiation in the presence or absence of 1,25-(OH)₂D₃ and Dex. The data shown represents the mean ±SD. (D) Results of TRAP staining of cocultured BM-derived osteoclast precursor cells and ALCAM^high cells, in which osteogenic differentiation was induced. TRAP-positive multinucleated cells are indicated by arrowheads. (a) no factor; (b) in the presence of 1,25-(OH)₂D₃ and Dex. Scale bar, 100 μm. (E) Total LMCs (a and d) ALCAM^low/− cells (b and e), and ALCAM^high cells (c and f) were induced to undergo adipogenic differentiation (as described in Materials and Methods) and stained with Nile red. (a–c) control medium; (d–f) MDI medium. Scale bar, 200 μm. (F) Alcian blue staining after chondrogenic induction. Total LMCs (a and d), ALCAM^low/− cells (b and e), and ALCAM^high cells (c and f) were induced to undergo chondrogenic differentiation as described in Materials and Methods. (a–c) no factor; (d–f) in the presence of BMP-2. Scale bar, 100 μm.

Figure 4. — Osteoblasts, adipocytes, and chondrocyte differentiation. (A) Total LMCs (a, d, and g), ALCAM^low/− cells (b, e, and h), and ALCAM^high cells (c, f, and i) were induced (d–i) or not-induced (a–c) to undergo osteogenic differentiation. After 2 wk, ALP staining was performed. Panels g–i show high power fields of d–f, respectively. Scale bars, 1 mm (a–f), 200 μm (g–i). (B) The region of alizarin red positive calcium deposition in ALCAM^high cells, in which osteogenic differentiation was induced for 2 wk. (a) no factor; (b and c) β-GP and ascorbic acid. (b) The center of culture plate. (c) The corner of the culture plate. (C) TRAP activities were measured on day 7 of cocultivation of BM derived osteoclast precursor cells and ALCAM^high cells, which were induced (hatched bar) or not induced (solid bar) to undergo osteogenic differentiation in the presence or absence of 1,25-(OH)₂D₃ and Dex. The data shown represents the mean ±SD. (D) Results of TRAP staining of cocultured BM-derived osteoclast precursor cells and ALCAM^high cells, in which osteogenic differentiation was induced. TRAP-positive multinucleated cells are indicated by arrowheads. (a) no factor; (b) in the presence of 1,25-(OH)₂D₃ and Dex. Scale bar, 100 μm. (E) Total LMCs (a and d) ALCAM^low/− cells (b and e), and ALCAM^high cells (c and f) were induced to undergo adipogenic differentiation (as described in Materials and Methods) and stained with Nile red. (a–c) control medium; (d–f) MDI medium. Scale bar, 200 μm. (F) Alcian blue staining after chondrogenic induction. Total LMCs (a and d), ALCAM^low/− cells (b and e), and ALCAM^high cells (c and f) were induced to undergo chondrogenic differentiation as described in Materials and Methods. (a–c) no factor; (d–f) in the presence of BMP-2. Scale bar, 100 μm.