Study Skeletal Muscle Of Microscopic Morphology Changes Of Rabbit Extremities After Sperfusing With 19% DMSO In Different Of Thawing And Freezing Methods

Objective:In this study,19%(v / v)DMSO perfusion rabbits were used to cut off the hind limbs.Four different cryopreservation methods were used to compare the degree of skeletal muscle injury in the composite tissues.To find the minimum methods of freezing and rewarming.Methods:Take only 25 healthy New Zealand white rabbits,which were randomly divided into control group(group A)and experimental group(group B?group C?group D and group E,B is slow-freezing and rapid-thawing,C is slow-freezing and slow –thawing,D is rapid-freezing and slow-thawing,E is rapid-freezing and rapid-thawing).All legs used in the experiment were broken at the hip joint.The control group was made directly samples after amputated limb.Experimental group in 19% dimethyl sulfoxide injection,heparin saline water elution,after dealing with different freezing and thawing groups.Skeletal muscle specimens were made in the same manner with the control group.Pathological sections were stained with HE to observe the degree of skeletal muscle injury.According to the results of pathological staining,the degree of damage to skeletal muscle tissue was analyzed statistically.Results:The pathological sections of skeletal muscle after HE staining were as following:the control group(group A):normal muscle cell and the nucleus form,no muscle fiber destruction and vacuolization occurred in skeletal muscle tissue;experiment group B:the cell of muscle is swelling,the muscle fiber is breaking,but the tissue morphology is still normal;Experimental group C:the edema of muscle cell is mild,with a small amount of vacuoles,the part of the muscle fibers is broken;Experimental group D:the edema of muscle cell is serious,with a small amount of vacuoles,the part of the muscle fibers is broken;Experimental group E:the edema of muscle cell is serious,a large number of vacuoles,the part of the muscle fiber is dissolved.Conclusions:1.There was a significant difference between the experimental group and the control group under the condition of 19% v / v dimethylsulfoxide perfusion,which is indicated that the cryopreservation of the cryopreserved cells had a significant difference between the experimental groups and the control group;2.The difference between the two groups was statistically significant,indicated that the degree of skeletal muscle injury due to different methods of freezing and rewarming difference;3.Experimental groups skeletal muscle injury from light to heavy:slow-freezing and rapid-thawing group <slow-freezing and slow-thawing group < rapid-freezing and slow-thawing group < rapid-freezing and rapid-thawing group.We can consider that the slow-freezing and rapid-thawing is the best methods of freezing and rewarming ways.