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Soft material-based microculture system having air permeable cover sheet for the protoplast culture of Nicotiana tabacum

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Abstract

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 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 cover glass and the PDMS sheet, respectively, and the microcolonies were formed well inside the PDMS covered chamber after 10 days.

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Abbreviations

BA:

Benzylaminopurine

FDA:

Fluorescein diacetate

MES:

2(N-Morpholino) ethanesulfonic acid

NAA:

1-Naphthalene acetic acid

PDMS:

Polydimethylsiloxane

REVs:

Relative evaporated volumes

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Acknowledgments

This study was supported by a grant of the Ecotechnopia 21 Project, Ministry of Environment, Republic of Korea (052-041-037).

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Authors and Affiliations

  1. Department of Biomedical Engineering, Korea University, Anam-dong Seongbuk-Gu, Seoul, 136-701, South Korea

    Jong Il Ju, Ju Yeoul Baek & Sang Hoon Lee

  2. Korea Artificial Organ Center, Korea University, Anam-dong Seongbuk-Gu, Seoul, 136-701, South Korea

    Jong Il Ju, Ju Yeoul Baek & Sang Hoon Lee

  3. Department of Medical Laser Graduate School,, Dankook University, San 29, Anseo-dong, Cheonan, Chungnam, 330-714, South Korea

    Jong Il Ju

  4. Department of Biology, # 419 The First science Hall, College of Advanced Science, Dankook University, 29 San Anseo-dong, Cheonan, 330-714, South Korea

    Jung-Moon Ko, So Hyeon Kim & Hyeon-Cheol Cha

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  1. Jong Il Ju
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  2. Jung-Moon Ko
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  3. So Hyeon Kim
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Correspondence to Sang Hoon Lee.

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Ju, J.I., Ko, JM., Kim, S.H. et al. Soft material-based microculture system having air permeable cover sheet for the protoplast culture of Nicotiana tabacum . Bioprocess Biosyst Eng 29, 163–168 (2006). https://doi.org/10.1007/s00449-006-0068-x

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  • DOI: https://doi.org/10.1007/s00449-006-0068-x

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