A Study Of Culturing Dorsal Root Ganglia Explants In Vitro For Axon Stretch Grown

The research of axon stretch grown has important significance for the study of peripheral nerve injury repair and neural interface technology which are based on nerve tissue engineering, but there is no in vitro culture system for the axon stretch grown. Therefore, a system of culturing Dorsal Root Ganglia explants in vitro for axon stretch grown has been studied in this thesis.Firstly, the evidences for consisting the system of dorsal root ganglia explants、Aclar films and the Extracellular matrix as the material for biological modification have been found. Secondly, the experiment of biological modification of Aclar films is designed to find the suitable modification method. Finally, the in vitro culture system is applied to the axon stretch equipment. The experiment of Aclar films modification:(1) Extract dorsal root ganglion explants from newborn SD rats;(2) Modify the Aclar films with PDL、PLL、Collagen-I、LN、PDL combined with LN、PDL combined with Collagen-I, respectively;(3)Culture the dorsal root ganglion explants on the six modified Aclar films, design the comparison experiment;(4)Observe and record the results. The experiment of axon stretch grown:(1)Design the axon stretch equipment;(2)Culture the Dorsal Root Ganglia explants with the in vitro system in the stretch culture room;(3)Start the axon stretch project after the axon grow to the stretch films;(4)Observe and record the results.The results of the experiment:Aclar films modification:(1)After 72 hours, the growth differentiation of axons of the PDL case are the best, the average length of axons in PDL case :1191.34 ? 227.41μ m; the density of axons in PDL case:0.1898 ? 0.052perμ m, the amount different between PDL case and PDL+Collagen-I case、PLL case、Collagen-I case、LN case are statistically significant(P<0.05);( 2) After 14 days, the photos of immunofluorescence staining show that PDL case has a wealth of axons and support cells, while the axons of the PDL+LN case get out of surface of films. The experiment of axon stretch grown: after stretch grown 71 hours, with the traction force the average grown speed of the stretched axons is 0.866 mm per hour; after 164 hours, the axons grow 6.124 mm.The conclusions:(1)The Aclar films modified with PDL is suitable for growth and differentiation of dorsal root ganglia axons.(2)the dorsal root ganglion in vitro culture system of using Aclar films as substrate, PDL as bioactivity modification material and culturing, can be used to study axon stretch grown.