Figure 5-4266.

A: Summary of tumorogenesis activities tests. The heterotopic bones were surgically removed from NSE-BMP4 transgenic mice and implanted subcutaneously in the back of NES-BMP transgenic mice or WT littermates. Cells extracted from heterotopic bones also subjected to soft agar colony analysis. There was no positive colony in soft agar colony analysis, and no growth of implants was observed in recipient mice either. B: Summary of sex-mismatched cell transplantation. Peritoneal cells and mononuclear cells were isolated from 2-month-old male donor NES-BMP transgenic mice (red lines) or male WT littermates (black lines), and were injected subcutaneously to the inner side of female left hind limb of 2-month-old NES-BMP4 transgenic mice or WT littermates. “no” indicated there was no heterotopic ossification within 2 weeks. “H.O.” indicated there was heterotopic ossification within 2 weeks. Note that only female NES-BMP4 transgenic mice receiving the male NES-BMP4 donor cells developed heterotopic ossification. C: Dual-color FISH experiment to follow the donor cells in recipient mice. a: Y-chromosome (Cy3), (b) X-chromosome (FITC), (c) DAPI staining, (d) merge of a–c frozen section of tissue with heterotopic ossification from recipient mice. Note the typical chondrocytes or hypertrophic chondrocytes morphology and nuclei staining, and more importantly, many of typical chondrocytes or hypertrophic chondrocytes had both X and Y chromosome staining.