Rapid Generation of Mature Hepatocyte-Like Cells from Human Induced Pluripotent Stem Cells by a Microfluidic Device
| Autor: | Yen-Ting Kuan, 管妍婷 |
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| Rok vydání: | 2013 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 101 Induced pluripotent stem cells (iPSCs) derived from the reprogramming of somatic fibroblasts are similar to embryonic stem cells (ESCs), having the potential to differentiate into all cell lineages of three germ layers in vitro. Thus, iPSCs may serve for a wide range of clinical applications, including cell transplantation, drug screening, and patient specific disease modeling. Previously, we have successfully established an efficient and rapid three-step protocol to generate hepatocyte-like cells from human iPSCs. To date, this is the most rapid differentiation protocol reported which allowed only 12 days to generate hepatocyte-like cells. However, previous studies have indicated that cells take up growth factors while secreting cytokines and wastes in the conventional method of plate culture. It can’t precisely control cellular behavior, including proliferation and differentiation into specific cell types. Therefore, we developed a microfluidic device that provides a stable platform for cells to grow and at the same time maintains a constant flow rate in the culture chamber. In this study, we applied this microfluidic device to compare the hepatic differentiation of human iPSCs from that grown in conventional culture dishes. The results showed a more rapid growth rate of human iPSCs from the microfluidic device compared to culture dishes. Quantitative PCR data revealed that using microfluidic device promoted expression of hepatic genes during hepatic differentiation of human iPSCs. Human iPSC-derived hepatocytes in microfluidic device also possessed the ability to store glycogen by PAS staining. Taken together, these results indicated that our microfluidic device not only accelerates the growth rate but also promotes efficiency of hepatic differentiation from human iPSCs. We expect that the results of this study can provide an effective platform tool for stem cell researches and clinical applications in the future. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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