Selective elimination, clump production and patterning of hiPSCs by light on PAG-polymer-functionalized substrates
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1
National Institute of Advanced Industrial Science and Technology, Biotechnology Research Institute for Drug Discovery, Japan
Introduction: Utility of human stem cells has been increasingly examined for drug assay or tissue engineering. For these purposes, automatable cell culture method is required to implement the pure and efficient propagation of those cells. In the culture of human pluripotent stem cells to maintain their undifferenciated state and viability stably, the cell should be treated in cell clumps avoiding discrete state which induces apoptosis in the subculture process, and be purified by removing the differenciated portion timely. To provide powerful tool to process cell culture systems on a culture substrate in such circumstances, we developed a method to kill the cultured cells selectively by using the combination of photo-acid-generating (PAG)-polymer-functionalized culture substrates and micro-patterned light irradiation[1]. Recently we found that the cell monolayer culture of the substrate can be cut by irradiation of line pattern. In this study, we examine the application of this photo-manipulation to the maintenance culture of human iPS cells (hiPSCs) (Fig.1).
Experimental and Results: We synthesized a poly(methylacrylate) functionalized with visible-light-responsive PAG group[2], and fabricated a photo-responsive culture substrate by spin-coating the polymer solution on polystyrene culture surface[1]. We cultured the colonies of hiPS cell line (201B7) on the substrate and irradiated light locally in the areas, where the cells were undifferentiated, using a PC-controlled micropattern irradiation system. After 2 hours, we observed that the cell viability was diminished only in the irradiated area (Fig.2). Also we demonstrated to produce cell clumps with te similar size from the grown colonies by the photo-induced cutting without critical damage. Together with the above purification method, this photo-manipulation was expected to provide powerful tools in automatizing the propagation of the human pluripotent stem cells.
This study was financially supported by JSPS KAKENHI (25282148).
References:
[1] Sumaru et al. Biotechnol. Bioeng., 110, 348-352 (2013)
[2] Shirai et al. Prog. Org. Coat., 64, 175–181(2009)
Keywords:
Automation,
Light,
stimuli-response,
Polymeric material
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Biomaterials and cellular signaling
Citation:
Sumaru
K,
Morishita
K,
Takagi
T,
Satoh
T and
Kanamori
T
(2016). Selective elimination, clump production and patterning of hiPSCs by light on PAG-polymer-functionalized substrates.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.02334
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.