Effect of cooling rate, cryoprotectant and holding time at different transfer temperatures on the survival of cryopreserved cell suspension culture (Puccinellia distans (L.) Parl.)

L. Heszky, Zsolt Jekkel, Abdel Hamid Ali

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Reflexed saltmarsh-grass suspension cultures produced by seed callus were frozen to the liquid nitrogen temperature. Cooling rates, cryoprotectants and holding times were taken as a function of transfer temperatures. The highest survival of cells (45%) was found at a freezing rate of 1°C min-1, without cryoprotectant treatments. The cryoprotectants (proline, dimethyl sulphoxide, glycerol), used at different concentrations and transfer temperatures, increased the survival rate. The maximum value was 78% at 12.5% (w/v) of proline with -30°C transfer temperature. Considerable improvement of viability (from 0% to 95%) among the 12.5 and 15.0% (v/v) dimethyl sulphoxide cryopreserved cells was achieved by holding them at - 20°C for 10-30 min before plunging into the liquid nitrogen. A 20 min holding time at 15.0% (v/v) glycerol level and - 30°C transfer temperature significantly enhanced the viability of the explants from 42% to 92%. Plants were successfully regenerated from cells cryopreserved with proline (w/v) and dimethyl sulfoxide (v/v) levels of 12.5 and 15.0%, respectively.

Original languageEnglish
Pages (from-to)217-226
Number of pages10
JournalPlant Cell, Tissue and Organ Culture
Volume21
Issue number3
DOIs
Publication statusPublished - Jun 1990

Fingerprint

Puccinellia distans
cryoprotectants
cell suspension culture
Suspensions
cooling
Cell Culture Techniques
dimethyl sulfoxide
Dimethyl Sulfoxide
Temperature
Proline
proline
temperature
Glycerol
glycerol
Nitrogen
viability
liquids
Bony Callus
nitrogen
Poaceae

Keywords

  • cooling rate
  • cryoprotectant
  • holding time
  • Puccinellia distans
  • suspension culture
  • transfer temperature

ASJC Scopus subject areas

  • Biotechnology
  • Plant Science

Cite this

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title = "Effect of cooling rate, cryoprotectant and holding time at different transfer temperatures on the survival of cryopreserved cell suspension culture (Puccinellia distans (L.) Parl.)",
abstract = "Reflexed saltmarsh-grass suspension cultures produced by seed callus were frozen to the liquid nitrogen temperature. Cooling rates, cryoprotectants and holding times were taken as a function of transfer temperatures. The highest survival of cells (45{\%}) was found at a freezing rate of 1°C min-1, without cryoprotectant treatments. The cryoprotectants (proline, dimethyl sulphoxide, glycerol), used at different concentrations and transfer temperatures, increased the survival rate. The maximum value was 78{\%} at 12.5{\%} (w/v) of proline with -30°C transfer temperature. Considerable improvement of viability (from 0{\%} to 95{\%}) among the 12.5 and 15.0{\%} (v/v) dimethyl sulphoxide cryopreserved cells was achieved by holding them at - 20°C for 10-30 min before plunging into the liquid nitrogen. A 20 min holding time at 15.0{\%} (v/v) glycerol level and - 30°C transfer temperature significantly enhanced the viability of the explants from 42{\%} to 92{\%}. Plants were successfully regenerated from cells cryopreserved with proline (w/v) and dimethyl sulfoxide (v/v) levels of 12.5 and 15.0{\%}, respectively.",
keywords = "cooling rate, cryoprotectant, holding time, Puccinellia distans, suspension culture, transfer temperature",
author = "L. Heszky and Zsolt Jekkel and Ali, {Abdel Hamid}",
year = "1990",
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T1 - Effect of cooling rate, cryoprotectant and holding time at different transfer temperatures on the survival of cryopreserved cell suspension culture (Puccinellia distans (L.) Parl.)

AU - Heszky, L.

AU - Jekkel, Zsolt

AU - Ali, Abdel Hamid

PY - 1990/6

Y1 - 1990/6

N2 - Reflexed saltmarsh-grass suspension cultures produced by seed callus were frozen to the liquid nitrogen temperature. Cooling rates, cryoprotectants and holding times were taken as a function of transfer temperatures. The highest survival of cells (45%) was found at a freezing rate of 1°C min-1, without cryoprotectant treatments. The cryoprotectants (proline, dimethyl sulphoxide, glycerol), used at different concentrations and transfer temperatures, increased the survival rate. The maximum value was 78% at 12.5% (w/v) of proline with -30°C transfer temperature. Considerable improvement of viability (from 0% to 95%) among the 12.5 and 15.0% (v/v) dimethyl sulphoxide cryopreserved cells was achieved by holding them at - 20°C for 10-30 min before plunging into the liquid nitrogen. A 20 min holding time at 15.0% (v/v) glycerol level and - 30°C transfer temperature significantly enhanced the viability of the explants from 42% to 92%. Plants were successfully regenerated from cells cryopreserved with proline (w/v) and dimethyl sulfoxide (v/v) levels of 12.5 and 15.0%, respectively.

AB - Reflexed saltmarsh-grass suspension cultures produced by seed callus were frozen to the liquid nitrogen temperature. Cooling rates, cryoprotectants and holding times were taken as a function of transfer temperatures. The highest survival of cells (45%) was found at a freezing rate of 1°C min-1, without cryoprotectant treatments. The cryoprotectants (proline, dimethyl sulphoxide, glycerol), used at different concentrations and transfer temperatures, increased the survival rate. The maximum value was 78% at 12.5% (w/v) of proline with -30°C transfer temperature. Considerable improvement of viability (from 0% to 95%) among the 12.5 and 15.0% (v/v) dimethyl sulphoxide cryopreserved cells was achieved by holding them at - 20°C for 10-30 min before plunging into the liquid nitrogen. A 20 min holding time at 15.0% (v/v) glycerol level and - 30°C transfer temperature significantly enhanced the viability of the explants from 42% to 92%. Plants were successfully regenerated from cells cryopreserved with proline (w/v) and dimethyl sulfoxide (v/v) levels of 12.5 and 15.0%, respectively.

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