Time-resolved NMR metabolomics of plant cells based on a microfluidic chip.
The plant secondary metabolism generates numerous compounds harbouring pharmaceutical activity. In plants, these compounds are typically formed by different and specialised cell types that have to interact constituting a metabolic process chain. This interactivity impedes biotechnological production of secondary compounds, because cell differentiation is suppressed under the conditions of a batch bio-fermenter. We present a novel strategy to address this limitation using a biomimetic approach, where we simulate the situation in a real tissue by a microfluidic chamber system, where plant cells can be integrated into a process flow. We show that walled cells of the plant model tobacco BY-2 can be successfully cultivated in this system and that physiological parameters (such as cell viability, mitotic index and division synchrony) can be preserved over several days. The microfluidic design allows to resolve dynamic changes of specific metabolites over different stages of culture development. These results serve as proof-of-principle that a microfluidic organisation of cultivated plant cells can mimic the metabolic flows in a real plant tissue.
Botanical Institute, Molecular Cell Biology, Karlsruhe Institute of Technology (KIT), Kaiserstr. 2, D-76131 Karlsruhe, Germany. Electronic address: Jan.Maisch@kit.edu.
Jan
J
Maisch
Institute of Microstructure Technology, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany.
Kristina
K
Kreppenhofer
Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, D-76131 Karlsruhe, Germany; Institute for Biological Interfaces, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany.
Silke
S
Büchler
Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, D-76131 Karlsruhe, Germany; Institute for Biological Interfaces, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany.
Christian
C
Merle
Institute of Microstructure Technology, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany.
Shukhrat
S
Sobich
Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, D-76131 Karlsruhe, Germany; Institute for Biological Interfaces, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany.
Benjamin
B
Görling
Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, D-76131 Karlsruhe, Germany; Institute for Biological Interfaces, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany.
Burkhard
B
Luy
Institute of Microstructure Technology, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany.
Ralf
R
Ahrens
Institute of Microstructure Technology, Karlsruhe Institute of Technology (KIT), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany.
Andreas E
AE
Guber
Botanical Institute, Molecular Cell Biology, Karlsruhe Institute of Technology (KIT), Kaiserstr. 2, D-76131 Karlsruhe, Germany.
Peter
P
Nick
D009682:Magnetic Resonance Spectroscopy; D055432:Metabolomics / Q000379:methods*; D044085:Microfluidics / Q000379:methods*; D010641:Phenotype; D059828:Plant Cells / Q000502:physiology; D013997:Time Factors; D014026:Nicotiana / Q000166:cytology
Microfluidics; NMR metabolomics; Tobacco cells (Nicotiana tabacum L. cv. BY-2)