Experimental Study Of The Directed Differentiation Of Neural Stem Cells And Transplantation In The Treatment Of Parkinson's Disease

Backgrounds and objectives Found of neural stem cell is the most important progress in neurobiology. It not only challenges the traditional un-regeneratable opinion, provide ideal vector of study on neuro-development and mechanism of neurogenesis, but also bring hope to the treatment of many neural diseases. Culture and expansion of NSC is the basement of the research. Objective of this part is to culture NSC in vitro, observe its biological property and make bases for further study on differentiation.Methods Using serum free culture medium and mitogen factors, NSC of rat and human fetal were isolated and cultured. Technique of long-term culture and single-cell clone culture was used to confirm the character of self-renewing. Ability of multi-directional differentiation and specific antigen, nestin, were detected by immunocytochemistry (ICC). BrdU was used to show cell stage in division. Effect of combination of EGF,bFGF on proliferation of rat NSC and LIF on human NSC were evaluated by clone forming and cytometry.Results The cultured NSC showed multipotential of differentiate into neurons and glial cells and the ability of self-renewing. Cells expressed nestin and many were active in division stage. The population of the NSC expanded quickly and the total number of the cell increased 2.4×104 folds after 15 times' passage. The property of NSC was stable after long culture and cry preservation. Combination of EGF and bFGF stimulated cell proliferation than used single. Addition of LIF in human NSC culture could promote cell proliferation and expansion. Conclusions The cultured NSC express nestin and have the ability of self-renewing and multi-directional differentiation. Combination of EGF and bFGF could promote the proliferation of rat NSC while LIF accelerate expansion of human NSC.