Chapter 34
Cryopreservation of Plant Cell Lines Using Alginate
Encapsulation
Heinz Martin Schumacher, Martina Westphal, and Elke Heine-Dobbernack
Abstract
Plant cell cultures consist of single cells or cell clusters growing as callus or suspension. Such cell cultures
may be able to produce secondary metabolites and/or possess embryogenic potential. Therefore, they can
be used for very different purposes in research, biotechnological applications, as well as for plant propagation. Cryopreservation is the only practical method to preserve such cultures until they are needed.
Different cryopreservation approaches that have been developed for differentiated plant tissues including
slow freezing, vitrification, and encapsulation/dehydration have also been applied to plant cell cultures.
The controlled rate or slow-freezing approach, however, remains to be the gold standard for cell cultures. In
this chapter, a standard slow-freezing cryopreservation procedure in combination with alginate immobilization is presented for long-term preservation of plant cell cultures.
Key words Suspension culture, Embryogenic, Dedifferentiated, Callus culture, Slow freezing, Controlled rate freezing, Immobilization, Alginate
1 Introduction
Plant cell cultures have a wide range of different applications
including secondary metabolite production [1], phytopharming
[2], the propagation of plants by embryogenic cell cultures [3],
and as model systems in fundamental research [4, 5]. For all of
these applications, reliable long-term preservation methods are
essential. So far, cryopreservation is the only possibility to realize
long-term preservation of plant cell cultures and has been established especially for biotechnological applications [6, 7].
A standard protocol for suspension cultures growing as single
cells or small cell clumps based on a controlled rate freezing
approach has been developed in 1980 by Withers and King
[8]. In recent years, several fundamentally different techniques,
including vitrification [9, 10] and encapsulation/dehydration
[11–13], have been developed for plant tissues, and many of
them have also been applied to cell suspension cultures. Most
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_34, © Springer Science+Business Media, LLC, part of Springer Nature 2021
639
Cryopreservation of Plant Cell Lines Using Alginate
Encapsulation
Heinz Martin Schumacher, Martina Westphal, and Elke Heine-Dobbernack
Abstract
Plant cell cultures consist of single cells or cell clusters growing as callus or suspension. Such cell cultures
may be able to produce secondary metabolites and/or possess embryogenic potential. Therefore, they can
be used for very different purposes in research, biotechnological applications, as well as for plant propagation. Cryopreservation is the only practical method to preserve such cultures until they are needed.
Different cryopreservation approaches that have been developed for differentiated plant tissues including
slow freezing, vitrification, and encapsulation/dehydration have also been applied to plant cell cultures.
The controlled rate or slow-freezing approach, however, remains to be the gold standard for cell cultures. In
this chapter, a standard slow-freezing cryopreservation procedure in combination with alginate immobilization is presented for long-term preservation of plant cell cultures.
Key words Suspension culture, Embryogenic, Dedifferentiated, Callus culture, Slow freezing, Controlled rate freezing, Immobilization, Alginate
1 Introduction
Plant cell cultures have a wide range of different applications
including secondary metabolite production [1], phytopharming
[2], the propagation of plants by embryogenic cell cultures [3],
and as model systems in fundamental research [4, 5]. For all of
these applications, reliable long-term preservation methods are
essential. So far, cryopreservation is the only possibility to realize
long-term preservation of plant cell cultures and has been established especially for biotechnological applications [6, 7].
A standard protocol for suspension cultures growing as single
cells or small cell clumps based on a controlled rate freezing
approach has been developed in 1980 by Withers and King
[8]. In recent years, several fundamentally different techniques,
including vitrification [9, 10] and encapsulation/dehydration
[11–13], have been developed for plant tissues, and many of
them have also been applied to cell suspension cultures. Most
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_34, © Springer Science+Business Media, LLC, part of Springer Nature 2021
639