| Summary: | 碩士 === 國立臺灣大學 === 動物科學技術學研究所 === 102 === According to the statistic from International Diabetes Federation in 2013, there is 7 billion people in the world and 28 million people suffering from type 1 diabetes (T1D) in 382 million diabetes patient. T1D is caused by an auto-immune reaction where the body’s immune system attacks the β-cells which could produce insulin to maintain blood glucose. Due to the developmental plasticity of mesenchymal stem cells (MSC), MSC-based therapeutic intervention has become a promising strategy to replace injured tissues. However, a major obstacle of applying MSC is required large amounts of MSC for transplantation. To tackle this issue, we seek for adipose-derived stem cells (ADSCs) owing to their ease of isolation, abundant sources and multipotency and would like to further enhance their proliferative ability and keep differentiation characteristics.
We isolated ADSCs from porcine back fat region of subcutaneous adipose tissues (pADSCs) and showed that these ADSCs expressed the MSC surface markers including CD29, CD44, CD90 and MHC I. These pADSCs also sustained the multipotency to differentiate into adipocytes, chondrocytes and osteocytes. We cultured pADSCs in basal medium supplemented with or without growth factors such as fibroblast growth factor 2 (FGF2) or fibroblast growth factor 4 (FGF4) to determine the proliferation by MTT analysis and cell counting. Our results demonstrated that both FGF2 and FGF4 increased the proliferation of pADSCs. In our experiments, FGF2 also increased ERK phosphorylation. Therefore we speculate that FGF2 may work on ERK pathway to promote pADSC proliferation. Furthermore, FGF-supplemented pADSCs could be induced to differentiate into insulin producing cells. Determined by immunofluorescent staining and examined by confocal microscopy, quantitative PCR and ELISA, β-cells markers insulin was highly expressed in FGF-supplemented pADSCs. These results indicated that FGF2 increases pADSCs proliferation without compromising the β-cells differentiation characteristics of pADSCs.
To conclude, in this study, we determined the optimal culture condition for pADSCs with either FGF2 or FGF4. These pre-conditioned pADSCs provide a potential strategy to increase pADSCs proliferation for transplantation in treating type 1 diabetes or other autoimmune diseases.
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