The Experiments Of The BMSC To The Nerve Cell Differentiation Supernatant After Induction To The Therapeutic Effect Of Mci-rats

Objective: Explore the Supernatant of three generations,four generations and fivegenerations of BMSC from rats to the nerve cell differentiation after 12h induction to thetherapeutic effect of mci-rats.Methods: 1.Segregation,culture and identification the BMSC. BMSC adopted the bone marrowto stick wall culture method, take the third generation of BMSC, adds CD29, CD44 and CD34antibody, the same type IgGI to contrast and comparison and the blank, detection by flowcytometric. 2. The induction and appraisal. Culture medium containing 0.2 volume fraction offetal bovine serum and 1 mmol/Lβ-mercapto-ethanol (β-ME) were used for 24-hourpre-induction and then take off the Culture medium and adds bFGF 20 ng/mL with 0.1 volumefraction of fetal bovine serum. Demonstrate by Immunocytochemistry. 3-month-old rats for tenis normal group to stand the standard by Morris water maz Hidden platform getexperiment,screen 40 ones from 15—22- month-old rats and Randomly divided into fourgroups. Normal saline, three generations,four generations and five generations of Supernatantinjected into the hippocampus respectively.After 4 weeks these 4 group of rats do Morris watermaz Hidden platform get experiment again. 4. CDK5 content measurement byImmunocytochemistry.Results:1. The bone marrow stromal cells can be induced to differentiate into NLCs effectively.Flow cytometer studies presented that the cultured BMSC were CD29、CD44positive and CD34negative,which displaying features of BMSC.2. Use culture medium containing 0.2 volume fraction of fetal bovine serum and 1mmol/Lβ-mercapto-ethanol (β-ME) were used for 24-hour pre-induction，A few cells isgradually to the nuclei direction contraction，when adds bFGF the change is moreapparent.Each group we can see some cells NSE positive, the positive rate was 42.50±6.40,onlya few cells GFAP positive.3. Before and after injection the Morris water maz Hidden platform get experiment Cruisein the location of the incubation period difference line statistical analysis. except to the threegenerations and the five generations have no difference between the groups，others was statistically significant（p<0.05）.the group of three generations and the five generations arebetter then the four generations ones.4. The expression of CDK5 in limbic system, Kruskal-Wallis test can think four groupsCDK5 expression in population distribution are not same or not all sam（e =32.046，P<0.001）,Wilcoxon rank sum test show that except to the three generations and the five generations haveno difference between the groups，others was statistically significant,and the CDK5 expressionof three generations and the five generations are lower than the four generations ones.Conclusion：1. The Supernatant of BMSC from rats to the nerve cell differentiation after 12hinductioned have the therapeutic effect to MCI rats2. the group of three generations and the five generations of The Supernatant of BMSC arebetter then the four generations to improve the state of MCI rats.