Poly(dimethylsiloxane)-Based Micro Chamber with Air Permeable Cover Sheet for the Protoplast of Nicotiana tabacum Culture.

In plant cell culture, the delivery of nutrition and gas (mainly oxygen) to the cells is the most important factor for viability. In this paper, we propose a poly(dimethylsiloxane) (PDMS)-based micro culture system that is designed to have good aeration. PDMS is known to have excellent air permeability, and through the experimental method, we investigated the relation between the degree of air delivery and the thickness of the PDMS sheet covering the culture chamber. We determined the proper thickness of the cover sheet, and cultured protoplasts of Nicotiana tabacum in a culture chamber covered with a PDMS sheet whose thickness 400 µm. The cells were successfully divided, and lived well inside the culture chamber for 10 days. In addition, protoplasts were cultured inside the culture chambers covered with the coverglass and the PDMS sheet respectively, and the microcolonies were formed well inside the PDMS covered chamber after 10 days. [ABSTRACT FROM AUTHOR]