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. 2009 Jan 21;18(7):955–968. doi: 10.1089/scd.2008.0310

FIG. 10.

FIG. 10.

Intramembranous bone formation in calvarial defects by the transplanted osteoblast-like cells derived from human embryonic stem cell–derived mesenchymal stem cells (hESC-MSCs) cultured in osteogenic medium. (A) FAC sorting of APC-conjugated alkaline phosphatase (ALP) and green fluorescent protein (GFP) of cells before transplantation (Gate R2). (B) ALP staining of the sorted ALP-positive cells (blue) counter-stained with neutral red (red) (20×). (C,E) Histological images of newly formed bone (NB) near the margins of the defect (H&E, 40×). (D) Staining of human-specific nuclear antigen (HuNu) and (F) GFP staining of osteocytes in new bone matrix (arrow). (G) Negative staining of HuNu mAb on the mature host bone. (H) HuNu staining of mouse femur. (I) Negative control of HuNu and (J) GFP staining with omission of primary antibodies. Magnification of images C–F was 40× and G–J was 20×.