| Recently, biological species are becoming endangered or extinct and numbers of the species have being quickly decreased due to more and more serious destruction of our environment, leading to deeply influences on existence and development of human being. Therefore, more and more attentions have been paid on the serious issue, and preservation techniques of biological species have been studied in developed countries. Among the them is the cryopreservation technique, which is becoming a hot topics in biological researches. Vitrification as a simple alternation method for cryopreservation has been successfully used for cryopreservation of plant shoot tips, embrogentic cells, protoplasts and suspension cultured-cell and so on.Vitrification is an new technique for cryopreservation of biological materials. A procedure was presented on cryopreservation by vitrification of suspension cultured-cells of Rhodiola fastigiata. It is showed that the survival rate of the suspension cultured-cells was influenced by the pre-culture, the loading, the dehydration, the freeze-thawing and the unloading treatments. An optimal procedure was obtained by a pre-culture of the cells for 1 hour in MS medium supplemented with 5% DMSO first. The cells were next loaded with 80% PVS2 for 40 min at room temperature and exposed to 100% PVS2 at 0°C for 50 min. The solution was then changed with fresh PVS2 before being plunged into liquid nitrogen. Unloading treatment of the cells after a period of freezing in liquid nitrogen, following a rapid warming at 40°C, was taken for three times (each time 10 min) with a 1.2 mol/L sucrose solusion. The survival rate of the suspension cultured-cells following the optima procedure was reached to 82.70% by TTC test.
|
PDF Full Text Request