Freezing Characteristics of Cultured Catharanthus roseus (L). G. Don Cells Treated with Dimethylsulfoxide and Sorbitol in Relation to Cryopreservation

Abstract

The freezing behavior of dimethylsulfoxide (DMSO) and sorbitol solutions and periwinkle (Catharanthus roseus) cells treated with DMSO and sorbitol alone and in combination was examined by nuclear magnetic resonance and differential thermal analysis. Incorporation of DMSO or sorbitol into the liquid growth medium had a significant effect in the temperature range for initiation to completion of ice crystallization. Compared to the control, less water crystallized at temperatures below -30 degrees C in DMSO-treated cells. Similar results were obtained with sorbitol-treated cells, except sorbitol had less effect on the amount of water crystallized at temperatures below -25 degrees C. There was a close association between the per cent unfrozen water at -40 degrees C and per cent cell survival after freezing for 1 hour in liquid nitrogen. It appears that, in periwinkle suspension cultures, the amount of liquid water at -40 degrees C is critical for a successful cryopreservation. The combination of DMSO and sorbitol was the most effective in preventing water from freezing. The results obtained may explain the cryoprotective properties of DMSO and sorbitol and why DMSO and sorbitol in combination are more effective as cryoprotectants than when used alone.

Applications of thermal analysis in cryopreservation of plant cells and organs

“…Vitrification of encapsulating material has also been obtained after incubation with vitrification solution containing sucrose and ethylene glycol. It seems that water contents of plant organs (shoot-tips or somatic embryos) that optimise survival in liquid nitrogen are in the range 20 and 30% [10,12,24].…”

Cell and tissue cultures ofCatharanthus roseus (L.) G. Don: a literature survey

“…Nevertheless, several successful procedures have been reported [217]. A protocol for C. roseus suspension cultures is presented in Table 12 [218][219][220]. The recovery of living cells was about 60% and they were able to produce indole alkaloids.…”

Impact of pr-10a overexpression on the cryopreservation success of Solanum tuberosum suspension cultures

“…Therefore, a substantial dehydration of cells is essential for their successful cryopreservation (Chetverikova 2008). This is usually achieved by the vitrification of plant cell material using PVS2 solution (Sakai et al 1990), the treatment with cryoprotectants such as dimethylsulfoxide (DMSO) (Chen et al 1984a) or a mixture of sucrose, glycerol and DMSO (DGS), as well as the addition of proline (Withers and King 1979) or carbohydrates such as sorbitol (Chen et al 1984b), trehalose (Bhandal et al 1985) or sucrose (Lu et al 2009). Unfortunately, increased dehydration results in more severe chemical and mechanical damages of the cells (Kaviani 2011).…”