Comparison of three techniques for cryopreservation and reestablishment of long-term Gentiana tibetica suspension culture.
Cryo Letters
; 27(5): 269-82, 2006.
Article
em En
| MEDLINE
| ID: mdl-17256062
ABSTRACT
Cryogenic storage of cell suspensions allows long-term maintenance of cultures. The main purpose of the study was to develop a successful cryogenic protocol for 10-year-old embryogenic cell suspensions of G. tibetica. We examined three techniques of freezing (I) controlled-rate cooling with various cryoprotectants (0.1-0.5 M DMSO, 0.5-1.0 M sucrose, 0.5-1.0 M glycerol, 0.25-1.0 M proline) or preculture with 0.4 M sorbitol and cryoprotectants (0.065-0.1 M DMSO, 0.2-0.8 M proline), (II) vitrification (PVS2) and (III) encapsulation. Cell viability was assessed by the TTC test and biomass increase. After controlled-rate cooling the majority of cells were lethally damaged, with only 3% viability observed. Vitrification and encapsulation approaches were more effective, assuring high levels of post-thaw viability ca. 85% and 7%, respectively. The encapsulation procedure gave faster recovery of the culture suspension than did vitrification, and ensured culture homogeneity and embryogenic competence.
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Base de dados:
MEDLINE
Assunto principal:
Sementes
/
Criopreservação
/
Gentiana
Idioma:
En
Ano de publicação:
2006
Tipo de documento:
Article