Sonic Hedgehog (Shh) is expressed in embryonic tissues that are critical for the patterning of the developing central nervous system, somite, and limb. It is also involved in whisker, hair, foregut, tooth, and bone development. Shh regulates neural and hematopoietic stem cell fate and is important for thymocyte differentiation and proliferation as well as T cell determination. In adult tissue Shh is associated with cancer development and tissue remodeling following injury. It has been reported that SHH can induce alkaline phosphatase production in C3H10T1/2 mouse embryonic fibroblast cells. To test the bioactivity of SHH, C3H10T1/2 cells were seeded into 24-well plate at a density of 1x105 cells/mL , and allowed to attach overnight before treated with certain concentrations of SHH for 24h and alkaline phosphatase levels in the cell supernatant were determined by ELISA. Alkaline phosphatase levels in the cell supernatant of C3H10T1/2 cells increased significantly after stimulated with SHH have shown in Figure1.