The Study Of Differentiation From Adipose-derived Stem Cells Of Mouseinto Neural-like Cells

Objective:To detect and fix the method of the acquirement and proliferation and passage of adipose derived stem cells., then prove its multiple-differentiation by inducing it into adipose cells and osteocytes.Methods:prepare two SD rats, about120g, no matter male or female. Firstly, the adipose tissue was obtained from the inguinal fat of the SD rats; Secondly, cut the adipose tissue and filtrate it by filter; Thirdly, put the medium containing ADSCs into a25cm2petri dish, then culture and passage. Fourthly, The third passage cells were identified by stem cells common marker CD34, CD44, CD45and CD106by Flow Cytometry. At last, change the basic medium by specific medium to induce ADSCs into adipose cells and osteocytes.Results:I can culture, proliferate and passage ADSCs successfully. The result of Flow Cytometry is as follows:CD34(+), CD44(+), CD45(-) and CD106(+), which prove the cells are ADSCs. Lastly, according to the oil red O staining and Alkaline phosphatase staining, we can see the adipose cells and osteocytes differentiated from ADSCs.Conclusion:There are ADSCs in adipose cells. And they can be obtained and cultured and proliferated successfully. They have the ability to differentiate into other cells, for example, adipose cells and osteocytes. Objective:To differentiate ADSCs into neuron-like cells using inducing medium.Methods:First of all, the ADSCs adhered on the flask were digested with trypsin when the cells reach80%confluency, and then there were about105cells adhered onto the6well plate. Secondly, the ADSCs in the6well plate were cultured using neurobasal medium containing B27supplement, bFGF and EGF. During differentiation,50%of the media was replaced every3days, and we also can saw the free floating neurospheres that were released from the cells culture surface in the media after5days. The neurospheres after7days were detected by immunocytochemistry using Nestin antibody. At last, spheres were cultured and maintained for10days in neurobasal medium containing only B27supplement, the detected the cells after differentiation by immunocytochemistry of NeuN,β-tubulin and GFAP.Results:Neural differentiation was analyzed through the detection of the expression of neuronalmarkers (MAP2ab, Neu N and β-tubulin), in addition to GFAP as a marker of astrocytes.Conclusion:ADSCs can be induced into neural-like cells after culturing in the special medium.