Study Of The Adipose Tissue-derived Stem Cells In Peripheral Nerve Recovery

Part 1:Study of Adipose tissue-derived stem cells of SD rats by serial explant after enzymatic predigestionObjective:To harvest Adipose tissue-derived stem cells(ADSCs) of SD rats by serial explant after enzymatic predigestion in vitro,in order to obtain a new cultivation method of ADSCs and compare the results with that from Zuk's method.Method:Adipose tissue was harvested from 1-month-old SD rat and divided into two groups.In group A,adipose was snipped to 8mm3 blocks and were put on a nylon net. After being digested by 0.25%trypsin 5 minuets and 0.1%collagenase typeⅠ20 minutes,the adipose blocks were cultivated for 2～3 more days and then moved into another culture flask.They were repeatedly cultured by serial explant method for three or four times.In group B,adipose tissue was treated by Zuk's method.Both groups were performed MTT and flow cytometry to study the culture results.Result:Cells of both groups had typical morphological characters of stem cells,and had simiar groth curves.They were positive in the ADSCs' marks CD29,CD44, CD54 and CD105,negative in CD34,CD45 and CD106.Group A has a higher positive rate than group B(P＞0.05).Conclusion:The method of harvest Adipose tissue-derived stem cells by enzymatic predigestion and serial explant in vitro is an alternative way for ADSCs cultivation, which has a similar result with that from the Zuk's.It can apply plenty of ADSCs samples in a short period of time,which enables its practical use in ADSCs cultivation in vitro. Part2:Study of the growth and neural differentiation of Adipose tissue-derived stem cells(ADSCs) co-cultured with SD rats Schwann CellsObjective:Investigate the function of Schwann cells on the growth and neural differentiation of Adipose tissue-derived stem cells(ADSCs) when they are co-cultured in the Transwell system.Compare the result with that from BME+BHA chemical culture methods.Methods:The ADSCs harvested in Step 1 and Schwann Cells harvested from the sciatic nerve of SD rats are divided into 3 groups:Group A,ADSCs and Schwann cells were cultured in Transwell co-culture system.Group B:ADSCs were induced by BME and BHA.Group C:contrast group.The morphology of ADSCs was observed and immunohistology was performed,the results were examed by statistics software. Results:ADSCs of Group A and Group B were partly differentiated to the cells which had long neurite and had a positive result in the NF-M staining but negative GFAP stain.Group A shows a great difference in neurite length compared with Group C (P＜0.05),and demonstrates a great difference in cell quantity with group B (P＜0.05)Conclusion:Schwann cells of SD rat had a neural differentiation inducing effect on ADSCs,which is similar to the BME+BHA culture method,and it had a better culture result in the cell quantity compared with group B,which enables its practical use in ADSCs neural differentiation.