Investigate The Therpation Effect On Transplantation Of FLK-1+Cells Derived From Mouse Fetal Liver In Mouse With The Model Of Liver Injuried By CCL4

Objective: Explore the features and functions of the fetal liver of FLK was-1+fetal liver cells to observe the efficacy of the fetal liver of FLK-1+was fetal livercell infusion for the treatment of carbon tetrachloride to build Kunming mice withacute liver injury model, and its role in repair of liver injury mechanism.Methods:Using immunomagnetic beads to extract the fetal liver of FLK-1+was cells. The stem cell markers before and after in vitro application of HGF andEGF induced cytokine induced by FACS before and after the fetal liver of FLK-1+was cell surface marker expression changes by RT-PCR detection ofcytokine-induced gene expression. Carbon tetrachloride as a liver poison, build acuteliver injury in Kunming mice and Kunming mice acute hepatic injury were randomlydivided into two groups: control group,20mice were tail vein enter the input salineFLK-1-positive cells; experimental group,20mice were input via the tail vein ofHGF and EGF-induced (1106cells). Blood determination of liver function after16hours, calculate the mortality at64hours, respectively, immediately after each groupof mice liver to remove fixed24h4°C to4%poly-formaldehyde solution, embeddedin paraffin and sliced, and to do routine HE staining to observe the pathologicalchanges in various organs. Using statistical software SPSS19.0to calculate andcompare the difference of two sets of indicators and mortality of liver function.Results:1. From17-19days of gestation C57BL/6mice in the single extractthe fetal liver cell suspension floating by trypan blue dye exclusion method, cellviability was over95%. Immunomagnetic bead method for the separation of fetalliver of FLK-1+was cells by flow cytometry analysis showed that the fetal liver ofFLK-1+was cell purity of more than98%.2. Very small number of fetal rat liver FLK-1+cells expressed albumin (positive rate0.6%) by96.38%of the HGF and EGF induced FLK-1+cells expressed albumin, thephenotypic characteristics of hepatocytes. In addition, RT-PCR detection of FLK-1+cells with high expression of Oct-3/4, Rex-1after5days, FLK-1did not express Oct-3/4and Rex-1expression was significantly induced by hepatocyte growth factordecreased or disappeared, suggesting that the loss of stem cell characteristics.3.99.9%carbon tetrachloride with peanut oil was diluted to0.4%,4%,40%threeworking concentration of10mice were each concentration group, biochemical testresults show: With the carbon tetrachloride concentration increased, AST, and ALT,TBIL index was significantly increased, there is a concentration-dependent manner (P<0.05); of ALB, the FIG decreased with increasing carbon tetrachloride concentration,there is also concentration-dependent, and tetrachloro-dioxide concentration (P<0.05). And only40%concentrations of the mice died,80%(8/10) mice were killedwithin16hours, of which four died within eight hours rest in paragraph8-16hours ofdeath.4. Fetal rat liver FLK-1+cells transplantation after16hours the control group, serumaspartate aminotransferase (AST,4095IU/L±1240IU/L) and alanineaminotransferase (ALT,3085IU/L,±1200IU/L, experimental group) weresignificantly higher than that (of AST1614IU/L±814IU/L, the ALT1714IU/L±990IU/L), but the control group, serum albumin (18.6±1.4) was significantly lowerthan the experimental group (27.0±3.2), the differences were statistically significant(p <0.05); no significant difference between control and experimental groups inserum total bilirubin, and fibrinogen (p>0.05). Mortality of about80%in the controlgroup, transplantation group, the mortality rate was61.5%, a slight decrease, but thedifference was not statistically significant (p>0.05).Conclusion:1. FLK-1can be used for rich poly fetal liver undifferentiated cells.2. Fetal rat liver FLK-1+cells with high expression of stem cell markers Oct-3/4,Rex-1induced by cytokines can be directed to differentiate into hepatocytes.3. Dfferent concentrations of CCl4can cause varying degrees of liver damage, while0.3mL/30g dose concentration of40%better simulate the pathophysiological changesin acute liver injury and good stability, is used to Kunming mice acute induced liverinjury model, a more ideal drugs.4. Fetal rat liver of FLK-1+was transplantation in a mouse model of drug-inducedliver injury have a therapeutic effect, can significantly improve their liver function, the mortality rate has improved to some extent.