The Comparative Study On The Methodology Of Separating The Epithelial Layer In Mice Bronchial Arch

The aim of this experiment is to find out the proper method of three kinds of classical methods ever used before ,which can separate the epithelial layer from the mice bronchial arch, for different purpose.The animal of this experiment is mice of KM in 9-30 somites period. We have adopted three methods, including the first method of the PBS solution containing 10mM EDTA adjusted to pH7.0 to hatch the first branchial arch, the second method of the D—hank's solution containing 0.25% trypsin adjusted to pH7.0 to hatch the first branchial arch and the third method of the D—hank's solution containing 0.125% trypsin and 0.005% EDTA adjusted to pH7.0 to hatch the first branchial arch. All the tests are undertaken in 20℃. Firstly, we get the proper time to separate the epithelial layer down by every method. Secondly, we separately deal with a embryo by every method for the proper time. Finally , we exam the results by pathological slide and special dye and scanning electronic microscope.By the first separating solution, hatched in 20℃ for 8-10 minutes, the epithelium detached from the arch above the basal membrane. The second separating solution is hard to operate. The third separating solution can separate the epithelium with some mesenchymal tissue from the arch for 5-7 minutes hatching in 20℃.The first method can be used for acquire exodermal epithelium from the bronchial arch without any mesenchymal tissue. The second method is not recommended. The third method is recommended for the purpose to get mesenchymal tissue without any epithelial components.