Soft material-based microculture system having air permeable cover sheet for the protoplast culture of Nicotiana tabacum.
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 polydimethylsiloxane (PDMS)-based microculture 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 having thickness of 400 microm. 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 cover glass and the PDMS sheet, respectively, and the microcolonies were formed well inside the PDMS covered chamber after 10 days.
Ju
Jong Il
JI
Department of Biomedical Engineering, Korea University, Anam-dong Seongbuk-Gu, Seoul 136-701, South Korea.
Ko
Jung-Moon
JM
Kim
So Hyeon
SH
Baek
Ju Yeoul
JY
Cha
Hyeon-Cheol
HC
Lee
Sang Hoon
SH
D000388:Air; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002478:Cells, Cultured; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D004548:Elasticity; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008422:Materials Testing; D008567:Membranes, Artificial; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods; D010539:Permeability; D011523:Protoplasts / Q000166:cytology / Q000502:physiology*; D012828:Silicones / Q000737:chemistry*; D014026:Nicotiana / Q000166:cytology / Q000254:growth & development*; D014462:Ultrafiltration / Q000295:instrumentation* / Q000379:methods
D004129:Dimethylpolysiloxanes; D008567:Membranes, Artificial; D012828:Silicones; C013830:baysilon