Effect of Two Different Cultural Methods on Osteoclast Bone Absorption Activity and Expression of ATPase a3 Gene / 浙江中医药大学学报

ABSTRACT

[Objective] To investigate the difference of bone Absorption function of the rat osteoclast(OC) in different cultural methods, and the difference in the levels of mRNA expression of bone Absorption key gene, ATPase a3 gene, and provide the basis for further investigation on the OC in vitro. [Methods] with mechanical separation method, mature osteoclast was separated from inner wall of 24 hours newborn rat long bones medullary cavity; and induction culture methods, bone marrow cell was induced to osteoclast like cell(OLC) by using 1,25(OH)2D3.The morphological and functional change of osteoclast was observed and the difference in the level of ATPase a3 mRNA expression was determined.[ Result]OC and OLC are multinucleated giant cells, which can be stained positive by potartrate resistant acid phosphatase(TRAP), and can form bone Absorption lacuna. The number of OLC in induction method is more than in mechanical separation method, but bone Absorption lacuna is smaller and shallower than the early stage of induction. The late stage of OCL is extremely similar to OC. There is no significant difference of ATPase a3 mRNA expression from the cells between mechanical separation 8 hour and induction culture 6 days, but either of them is significantly less than inducing 8 days. [Conclusion]Induction method can produce a large number of OLC which is superior to mechanical separation method, but its bone resorption function is weaker in the earlier stage, because bone resorption function is associated with the number of nucleus. The late stage of OCL is extremely similar to OC of mechanical separation method. And it can be used in any experiments.