Determination Of Cytotoxicity Of Fibrin Glue On Human Adipose Derived Stem Cells By LDH

Objective:In recent years,with fat stem cells(adipose derived stem cells,ASCs)findings,based on the principle of tissue regeneration is adipose tissue engineering got great development,adipose stem cells as seed cells in adipose tissue engineering research hotspot.In adipose tissue engineering construction,scaffold materials is the key part of the scaffold materials of tissue engineering to satisfy multiple conditions: have good biocompatibility,can be degraded in a specific time,etc.Fibrin glue(fibringel)is to simulate coagulation reaction,fibrinogen in under the action of thrombin formation has certain physical strength of jelly.There have been reports fibrin gel as tissue engineering scaffold material used in a variety of studies,however,fibrin gel as tissue engineering scaffold material composite ASCs is used to build fat is still lack of enough research.This research will detect fibrin glue for ASCs cytotoxicity,further enrich using fibrin glue to make theoretical achievements of adipose tissue holder.Methods:1,extracting fat source of stem cells,cultivation,take the third generation of cells inoculated into 96-well plates,parallel microscopic observation.Flow cytometry detection ASCs surface antigen and ASCs induced into fat and oil red O staining.2,the fibrinogen and thrombin mixed in different proportions,made of different concentrations of fibrin glue soakage extract.And divided into the following four groups: negative control group,50% fibrin glue group leaching solution,100% fibrin glue leaching solution group,positive control group(maximum enzymes release control).The LDH were used to detect the kits each medium,the content of LDH.3,24 h and 72 h two time points 50% fibrin glue leaching solution group,100%fibrin glue leaching solution,respectively,compared with negative control group,the data to mean + /-standard deviation,to input all data into the software for processing in the analysis.Through the single factor analysis of variance method comparison between groups,through the inspection method for comparison between groups,demonstrated significant difference.Results:1,optical lens,the first three generations ASCs for typical fibroblast sample shape,is long and thin spindle,have thick branches,nucleoli.2,flow cytometry detection surface antigen CD49 d ASCs,CD105 high expression of CD34,CD45,CD106 don't express or low expression.3,ASCs in a concomitant induction medium oil red O staining positive after two weeks.4,in 24 h and 72 h two time points 50% fibrin glue leaching solution group,100% fibrin glue leaching solution,respectively,compared with negative control group,the data to mean + /-standard deviation description,to input all data into the software for processing in the analysis.Compared between groups by single factor analysis of variance method,through testing method to carry on the contrast between two groups.Between the calculation results show that each group and control group had no significant difference(P > 0.05),prompted fibrin glue did not increase the leaching solution of ASCs LDH release,fibrin glue for ASCs no significant cytotoxicity.Conclusion:Different concentrations of fibrin glue on fat source of stem cells has no obvious toxicity,fibrin glue is expected to become one source of stem cells to grow fat scaffold material.