Developement of Microfluidic-Based Mitochondrial Delivering System for Therapy of Suspending Stem Cell( I )

We are going to develop a massive cell hole opening and large cargo delivering system for suspension-type cells that is going to deliver mitochondria into stem cells for cell therapy. With becoming older and older, the mitochondrial gene mutation of adult people are dramatically increasing. When doing autologous stem cell therapy, it may still exists the mitochondrial-related diseases due to stem cells’ mitochondrial DNA (mtDNA) mutation. Another interested topic, induced pluripotent stem cell (iPSC), is to induce somatic cell become stem cell. However, during programming iPS process, the mitochondria in cells keep fusion and fission, during which mtDNA is lack of protection mechanism, mtDNA is very easy to mutate. Since mtDNA editing is not as simplify as DNA in nuclear. Recently, one way is to modify specific peptide on cell membrane to let mitochondria transparent easily. The collected normal mitochondria from other health cells are implanted into target cells. Normally, this method need to culture cells in vitro, and may not be suitable for stem cell e.g. Mesenchymal Stem Cell (MSC). When we will use MSC for autologous stem cell therapy, because after in vitro culture, MSC will dramatically decrease their therapy ability resulting from mciro-environmental changed. In our goal, we will develop an active mitochondria deliver system that can achieve high throughput, parallel, and high efficient mitochondria deliver without in vitro culture. We will demonstrate the system in three ways. Firstly, we demonstrate the delivering efficiency by targeting normal cell. After delivery, we investigate cell metabolism to show healing ability after therapy. Secondly, we study stem cell, targeting to bone marrow MSC (BM-MSC). We investigate the viability and metabolism of mitochondria-transferred MSC. Furthermore, study their viability and metabolism when they differentiate to downstream cells. Thirdly, we focus on iPSC, and study the mitochondria delivery. We collect somatic cells from adult people and program iPS to become stem cells. Further, investigate gene mutation of iPSC before/after programming iPS, and see which showed less mutation and less mitochondrial defect. We also study their differentiation to see the cell metabolism of their downstream cell. We also study cell metabolism of differentiated iPS-MSC after delivering mitochondria in vitro. Finally, we study the stem cell healing ability and compare to BM-MSC and iPS-MSC.