Source
Commissariat à l'Energie Atomique, institut de Recherche en Technologies et Sciences pour le Vivant, Laboratoire de Physiologie Cellulaire Végétale, Unité Mixte de Recherche 5168 CNRS, UJF, INRA, CEA, F-38054 Grenoble, France. rbligny@cea.fr.
Abstract
ABSTRACT:
BACKGROUND:
The repeated weekly subculture of plant cell suspension is labour intensive and increases the risk of variation from parental cells lines. Most of the procedures to preserve cultures are based on controlled freezing/thawing and storage in liquid nitrogen. However, cells viability after unfreezing is uncertain. The long-term storage and regeneration of plant cell cultures remains a priority.
RESULTS:
Sycamore (Acer pseudoplatanus) and Arabidopsis cell were preserved over six months as suspensions cultures in a phosphate-free nutrient medium at 5°C. The cell recovery monitored via gas exchange measurements and metabolic profiling using in vitro and in vivo 13C- and 31P-NMR took a couple of hours, and cell growth restarted without appreciable delay. No measurable cell death was observed.
CONCLUSION:
We provide a simple method to preserve physiologically homogenous plant cell cultures without subculture over several months. The protocol based on the blockage of cell growth and low culture temperature is robust for heterotrophic and semi-autotrophic cells and should be adjustable to cell lines other than those utilised in this study. It requires no specialized equipment and is suitable for routine laboratory use.