Tobacco protoplast culture in a polydimethylsiloxane-based microfluidic channel

Abstract

Several advances have been made in the use of microfluidic devices for insect and mammalian cell cultures, but no reports of their use for plant cell cultures have been published. We, therefore, conducted a plant cell culture in a microfluidic device using polydimethylsiloxane. Nicotiana tabacum protoplasts were cultured in a variously shaped polydimethylsiloxane channel containing Nitsch medium supplemented with 0.5 g of NLN-13 vitamin mixture, 2.0 mg of alpha-naphthaleneacetic acid, and 0.5 mg of 6-benzyladenine per liter and 9% mannitol. Protoplasts in the polydimethylsiloxane channel showed cell division and microcolony formation within 4 weeks. The use of a microfluidic channel is a novel technique in the field of plant cell culture. The results of this study will encourage the utilization of polydimethylsiloxane-based microfluidic devices in plant cell engineering and cell analysis.

Time-resolved NMR metabolomics of plant cells based on a microfluidic chip

“…The reason is that these cells are not attached to a surface as mammalian cells, but have to be cultivated in suspension. To our knowledge, only two reports have been published, where plant cells were cultivated in a microfluidic chip (Ko et al, 2006;Wu et al, 2011). However, these were protoplasts, where the cell wall was removed and therefore highly distorted in their physiology.…”

Culture and chemical-induced fusion of tobacco mesophyll protoplasts in a microfluidic device

“…The increase in cell volume as well as the change in the arrangement of chloroplasts preceded cell division, which corresponded to the previous study on in vitro protoplast culture in a suspension in Petri dishes (Nagata and Takebe 1971;Coutts and Grout 1975). The Nagata and Takebe (1971) Petri dish 3 5-7 10 -- Nagata and Takebe (1970) Petri dish 2-4 4-6 6-7 [7 - Thomas and Rose (1983) Petri dish 2 ---14 Ko et al (2006) results showed that cell division of the protoplasts in the chamber is evident beginning on the third day of culture. The chloroplasts in the parent cell were averagely distributed into two daughter cells (Fig.…”

Plant Cells Use Auxin Efflux to Explore Geometry

“…However, these approaches remained merely technical and did not integrate cells. There are two reports, where plant cells- protoplasts from tobacco leaves- have been kept alive in a microfluidic channel over several days1617 – however, in both cases, cells were floating freely in a large compartment of around 1 mm width without any possibility to administer directional cues to the regenerating protoplasts. In contrast, the system described in the current work successfully integrates living, developing plant cells into a technical realization of a tissue, where either mechanical or chemical cues can convey directional information.…”