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The Effects of Different Cryoprotectants on the Cryo preservation of Rabbit Embryonic Stem Cells YUAN Xiao-hua, WANG Shu-fen, YANG Shi-hua, LI Ya-hui, CHEN Hong-wei, JI Wei-zhi Abstract An effective freezing-thawing technique is crucial for the clinical application of newly derived rabbit embryonic stem (ES) cells. The aim of this study was try to find an optimal cryopreservation protocol for rabbit embryonic stem cells using slow freezing-rapid thawing without a programmable freezer. We tested the effects of the following cryoprotective agents (CPAs) on the post-thaw survival, proliferation and differentiation capacity of rabbit embryonic stem cells: ethylene glycol (EG), dimethyl sulphoxide (Me2SO, DMSO), trehalose and glutamine. Trypan blue exclusion tests showed that, among the CPA treatments in this study, EG was more toxic to rabbit embryonic stem cells than DMSO. The highest survival rate (83.7%) was obtained when the rabbit embryonic stem cells was cryopreserved with 10%DMSO and 20 mmol/L glutamine in the ES cell culture media. Thawed ES cells kept their pluripotency and differentiation potential. Keywords Rabbit embryonic stem cells; Slow-rate cooling; Cryoprotectants; Glutamine

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