Study Of Viability And Post-transplantation Survival Rate Of Adipose Tissue Cryopreserved At Different Conditions

Objective To explore an appropriate cryoprotector for cryopreservation of adipose tissue under different temperatures. To study the viability and post-transplantation survival rate of frozen adipose tissue. Methods 1. Treated with different cryoprotectors, fat granules was stored at -16℃ (domestic refrigerator), -80℃ (ultra-low-temperature refrigerator) and -196 ℃ (liquid nitrogen). 24 hours later, the activity of creatine kinase was measured to screen the appropriate cryoprotector. 2. Treated with the appropriate cryoprotector, fat granules was cryopreserved at the different temperatures described above. Thawing at 2 weeks, 4 weeks and 8weeks later post-preserve respectively, the activity of frozen fat was deteced by determination of creatine kinase and trypan-blue staining method; the post -transplantation survival rate was studied by biological implantation. Grafts were dissected out 7 weeks later post injection, then its' volume was measured and Sudan III staining was performed. Results 1. At -196℃, while cryopretector was 15% dimethyl sulphoxide and 6% propanediol, the viability of adipose tissue was the best. At -80℃, the highest viability was obtained with the use of 15% dimethyl sulphoxide. It was 5% dimethyl sulphoxide ,3% propanediol and 1% polyethylene at-l6℃.2. After 8 weeks frozen storage, compared with that of fresh fat, the viability of adipose tissue was kept well at -196°C and -80℃group ,which reached to 70%...