| Bovine artificial insemination technology had been widely used for livestock production, associated with the production of semen freezing technology.Compared with the bovine sperm, pig sperm was more sensitive to temperature change. As the sperm cell membrane resulted in higher levels of unsaturated fatty acids during frozen thawed sperm function vulnerable to injury, after thawing, the survival rate of the sperm and the reproductive rate after artificial insemination were both decreased, It was made difficult to promote the semen cryopreservation technology.Glycerol was a conventional semen cryopreservation, with penetration into the cell membrane,could reduce cell water content, prevented formation of ice crystals within cells, but the glycerin in the protection of sperm cells due to penetration into the cell membrane resulted in expansion of damage within a hypotonic environment, not were used in semen cryopreservation in large-scale. the sperm could be provided energy by Metabolism of carbohydrates. it would not enter the cell, so that it could be increased the extracellular osmotic pressure, the freezing process, to set the intracellular to extracellular water seepage, causing cell dehydration and shrinkage, reduction of cell within the ice crystal formation, infiltration through its own characteristics as a low-temperature protective agent against extracellular role to protect cells from damage, it was important to cryoprotectants, but over the years of study, results be showed that the carbohydrate effect of freezing vitality of Pig Sperm is limited. Therefore the exploration and selection of different cryoprotectants and optimization of freezing procedures were the main direction of research in recent years.This study was diluted in freezing medium with different concentrations of Polysaccharides, LBP and laminaria, adding different concentrations of glycerol, the two factors in the cross-design conditions, the detection of pig sperm after thawing, survival rate, mitochondrial activity, the rate of plasma membrane integrity and acrosome integrity rate were studied at different concentrations of glycerol and the compatibility between the effect of polysaccharide obtained the following results:In a process of porcine semen freezing, in, the 0.25-1.25mg/mL Radix Phodiolae Polyose were added, based on the traditional dilution TCG.,frozen-thawed sperm motility were 44.4%(1.00mg/ml, P<0.05). The Integrity rate of cytolemma were 40.58%和40.04% (1.00 mg/ml and1.25mg/ml, P<0.05).The activity of mitochondria were 43.12% (1.00mg/ml, P<0.05);But in the Integrity rate of acrosome,all treatment group were no significant difference.In a process of porcine semen freezing, in, the 0.25-1.25mg/mL LMP were added, based on the traditional dilution TCG. frozen-thawed sperm motility were 44.4% and 40.77% (0.75mg/ml and 1mg/ml,P<0.05). The Integrity rate of cytolemma were 40.72%,40.03% and 40.00%(0.75mg/ml, 1mg/ml and1.25mg/ml, P<0.05).The activity of mitochondria were 43.12%(1.00mg/ml, P<0.05); The Integrity rate of acrosome were 59.29%和and 58.82% (1mg/ml and 1.25mg/ml, P<0.05),all treatment group were no significant difference.But the activity of mitochondria were no significant difference.3 In a process of porcine semen freezing, in, the 0.25-1.25mg/mL lamarin were added, based on the traditional dilution TCG. frozen-thawed sperm motility were:44.39% (0.25mg/ml, P<0.05);The different quality score were no significant difference to frozen sperm plasma membrane integrity rate and acrosome integrity rate. The activity of mitochondria were increased significantly by high density of lamarin (1.25mg/ml, P<0.05)The results show that different polysaccharides on the porcine sperm cryopreservation had different effects, their sperm cryopreservation results were Polysaccharides> laminarin> LBP,compared with motility rate, the integrity of plasmic membrane, the integrity of acrosome, the integrity of mitochondria.The two factors crossover design were carried out by using different concentrations of Polysaccharides, LBP, Laminaria all which were added to different concentrations of glycerol, result showed that polysaccharides and glycerol had combined effect, which 1.00mg/mL Radix Phodiolae Polyose and 2.5% glycerol were combined,0.75mg/mL LBP and 2.5% glycerol were combined,0.5mg/mL laminarin and 2.5% glycerol were combined,the frozen effect significantly increased compared with the control group, results showed that the added amount of glycerol were reduced through the addition of polysaccharides, could not be instead of all the glycerin, the sperm cell morphology and cell function were protected by interacted with the glycerol and the polysaccharides; The frozen sperm quality were increased by the interaction with the glycerol and the polysaccharides, but not a substitute for glycerol as frost protection agents.
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