BENEFICIAL EFFECTS OF PLURONIC F-68 AND ARTIFICIAL OXYGEN CARRIERS ON THE POST-THAW RECOVERY OF CRYOPRESERVED PLANT CELLS

Figures & data

Table I. Some Advantages and Disadvantages of the Cryopreservation of Cells

Advantages Storage and conservation of elite and endangered germplasms Facilitates germplasm transportation and distribution

Disadvantages Not applicable to all cell types Dependant on specific pre- and post-thaw treatments Cell loss on thawing Post-thaw physiological changes

Table II. A Typical Cryopreservation Protocol for Eukaryotic Cells

Pre-freeze treatment(s) Addition of cryoprotectant (e.g. dimethyl sulphoxide, glycerol) Freezing in liquid nitrogen ♦ ‘flash’ immersion ♦ programmed freezing Cryostorage Thawing Post-thaw treatment(s)

Table III. General Chemical and Physical Properties of Pluronic® F-68

Average values for a, b and c = 75, 30 and 75, respectively.
Mean molecular weight	8350
Melting point (°C)	50
Physical form at 20°C	solid
Cloud point (°C)^*	>100
HLB^**	29

^* Cloud point = temperature at which solutions of non-ionic surfactants become cloudy and undergo phase separation to yield a detergent-rich layer and an aqueous layer.

^** HLB = hydrophilic-lipophilic balance number, an empirical measure of the emulsifying power of a surfactant. Compounds with low HLB values (<20) are hydrophobic, whereas those with higher values (>20) are hydrophilic.

Data from References [43-44].

Table IV. Benefits of Using PFCs in Cell Culture Systems

Chemically and biologically inert Easily sterilised (e.g. by autoclaving) Recoverable and recycleable High respiratory gas solubility Scavengers of gaseous cellular products (e.g. ethylene) Provide a two-phase (PFC liquid–aqueous medium) interface and physical support system

From Reference [[60]].

Table V. Properties of Erythrogen™

Total haemoglobin content (g 100 ml^−1)	10.3
P50 (Torr at 37°C)	38.0
Methaemoglobin content (%)	< 4
pH	7.42
Stable ca. 1 year at 5–10°C

Lowe K. C., Davey M. R., Power J. B. Perfluorochemicals: their applications and benefits to cell culture. Trends Biotechnol. 1998; 16: 272–277

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