Abstract
With increasing interest in induced pluripotent stem cells (iPSCs) in the field of stem cell research, highly efficient infection of somatic cells with virus factors is gaining importance. This paper presents a method of employing microfluidic devices for dynamic cell culture and virus infection in a microchannel. The closed space in the microchannel provided a better environment for viruses to diffuse and contact cell surfaces to infect cells. The microfluidic devices were fabricated by photolithography and soft lithography. NIH/3T3 fibroblast cells were cultured in the microfluidic device in static and dynamic conditions and compared with the conventional culture method of using Petri dishes. Virus infection was evaluated using an enhanced green fluorescent protein virus as a model. Dynamic culture in the microchannel showed similar growth of cells to that in Petri dish culture, but the virus infection efficiency was four-times higher. The proposed dynamic culture system could be useful in iPSC research by providing efficient virus infection tools.
Highlights
The first demonstration of the reprogramming of mouse fibroblasts into induced pluripotent stem cells by Takahashi et al in 2006 triggered a major paradigm change in the field of stem cell research [1]
Because induced pluripotent stem cells (iPSCs) are derived from somatic cells and not from an embryo, they could solve an ethical problem in stem cell research, opening the possibilities of personalized stem cell therapy and disease models
EirfufescitnoffeMctiiocrnotfleusitdsiwc CerueltpuerrefoSrymstemd foonllVowiruinsgInthfeecteixopnerimental protocols described in Section 2.2, and the virus infection efficiency was examined at the three cell culture conditions
Summary
The first demonstration of the reprogramming of mouse fibroblasts into induced pluripotent stem cells (iPSCs) by Takahashi et al in 2006 triggered a major paradigm change in the field of stem cell research [1]. During the derivation process of iPSCs, somatic cells are infected with four factors, namely Oct[4], SOX2, KLF4 and c-MYC [2,3]. The low efficiency of infection with virus factors results in very low reprogramming efficiency. Microstencils enabled localized culture of cells in microscale patterns to allow subsequent analysis in engineered manners [10,11].
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