| Summary: | 碩士 === 國立陽明大學 === 口腔生物研究所 === 91 === 暫缺The term “stem cell” means a class of cells, which can self-renew and differentiate into different kinds of cells. The techniques of inducing stem cells to differentiate into all kinds of cells can be applied to repair damaged tissues. Adipose tissue derived stromal stem cell shares the same origin with bone marrow stromal cell, and also has the multi-potential of differentiation like bone marrow stromal cell. However, to acquire adipose tissue derived stromal stem cell is much easier, which makes it become a great replacement of bone stromal stem cell in the application of damaged tissue repairing. Our laboratory has already developed the technique of inducing dental papilla cell to differentiate into neuronal cells previously. On this base we believe that the adipose tissue derived stromal stem cell can be a better source than bone marrow stromal stem cells if the technique can be applied to adipose tissue derived stromal stem cell. Thus the main goal in this research is to apply this technique to study adipose tissue derived stromal stem cell.
In this research, adipose tissues from adult male S. D. rat abdomens are digested by enzyme, and then the low-density adipocytes are eliminated by centrifugal. Isolated adipose tissue derived stromal stem cells are cultured in Dulbecco's modified eagle media and FBS. The morphology of these adipose tissue derived stromal stem cells is recorded by reversed phase contract microscope. When dexamethasone, beta-glycerophosphate, and 2-phosphate ascorbate are added to culture media, those cells differentiate into calcifying cells.
Immunohistochemistry results show that cultured adipose tissue derived stromal stem cells express neurotrophin receptor Trk and P75NTR. Moreover, when nerve growth factor is added to the culture media of adipose tissue derived stromal stem cells, although no obvious morphology change is observed, the decrease of neuronal cell marker nestin, the increase of neuron maker NeuN, neurofilaments 200KD, and the increase of astrocyte maker GFAP are observed from the results of immunohistochemistry. There is no obvious change in the expression of oligodendrocyte maker A2B5. Infer from these results we think that the differentiation fate of nerve growth factor treated adipose tissue derived stromal cell has the inclination for neuron of astrocyte but not oligodendrocyte, and these cells are not terminally differentiated.
In summary, the results of this research showed that adipose tissue derived stromal cell has the potential to differentiate into neural cells. The method of purification those multi-potential stem cells and the way to induce neuronal differentiation can direct the focuses of future studies.
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