| Summary: | 碩士 === 國立成功大學 === 口腔醫學研究所 === 98 === Induced pluripotent stem (iPS) cells, which show similar functions to embryonic stem cells, can now be derived from human adult somatic tissues. Human embryonic stem cells research involves many ethical issues all the time, and the production of iPS cells now can avoid these issues. Recent researches have shown that introduction of Oct3/4, Sox2, c-Myc, and Klf4 into human fibroblast can reprogram human somatic cells, and generate ES-like pluripotent stem cells. In the previous studies of cancer stem cells in our laboratory, we found human normal oral mucosa tissues expressed Oct3/4 based on immunohistochemistry (IHC). In this research, we want to further confirm that whether human oral mucosa tissues can indeed produce Oct3/4, even more other defined factors, to study if oral mucosa fibroblast (OMF) is a good source to produce iPS cells.
At first, we analyzed oral mucosa tissues by IHC and found that oral mucosa tissues expressed Oct3/4 and Sox2 by IHC. Then we used cell culture to further study whether oral mucosa fibroblast also expressed Oct3/4 and Sox2. We collected cell lysate from oral mucosa fibroblast to perform Western blotting and found OMF didn’t express Oct3/4 and Sox2 distinctly. Even so, we considered that it’s easy to get the oral cells. We hypothesized that if we can produce iPS cells by oral mucosa fibroblast successfully, oral mucosa fibroblast are still a good source to produce iPS cells. We tried to produce lentivirus to introduce Oct3/4, Sox2, c-Myc, and Klf4 into oral fibroblast. We expected to produce iPS successfully by oral cells and it will be useful to study disease therapy.
On the other hand, we used mouse iPS cells, which we bought from Japan, to study the application of iPS cells on disease therapy. We tried to induce the mouse iPS cells, which expressed green fluorescence protein (GFP), to differentiate into osteoblast. Then we supplied the osteoblast to cranial osseous defect on mice by HA-TCP to enhance the regeneration of bony defect. We analyzed the repair of bony tissue by IHC, and we stained the bony tissue with green fluorescent protein to confirm these cells were differentiated by iPS cells which expressed green fluorescent protein. In addition, we also studied the distribution of these cells in the animal model by observing the position of the green fluorescence by using IVIS (in vivo imaging system). By this research, we’ll further understand the application of iPS cells on oral and maxillofacial surgery therapy.
|
|---|
