The Effects Of Rats Islet Cells In Vitrification Cryopreservation

Objective To investigate the effects of rats islet cells in vitrification cryopreservation and building islet cells bank by the way.To observe the effects of rats islet cells in cryoprotectant according to the joint application of different ratios.Choose better program and investigate the mechanisms.Methods Islet cells of isolation and purification are divided into four groups.The purity is qualified(＞60%).A group(control group) fresh islet cells.Observe the difference rewarming islet cells with freh islet cells.Group B 10%(V/V) DMSO+10%(V/V) EG. Group C10%(V/V) DMSO+10%(V/V) EG+10%(V/V) Formamide,Group D10% (V/V) DMSO+20%(V/V) EG+10%(V/V) Formamide.B,C and D Groups are preserved by vitrification cryopreservation.Mixing islet cells with cryoprotectant,directly into liquid nitrogen,37℃rapid rewarming after 7 day and 14 day respectively.Different concentrations of sucrose solution,Hanks exgraft the cryoprotectant classify.RPMI-1640 cultures islet cells, containing 10%(v/v) fetal bovine serum.Put the culture bottles in 95%O₂,5%CO₂ incubator at 37℃for overnight.Through AO-PI staining,calculate the staining cells in fluorescent microscope and calculate the survival rate of islet cells,the survival rate indirectly reflects the cellular structure integrity.Test the insulin secretion reflects the islet cells function.Comprehend two indexes and evaluate vitrification cryopreservation.Results(1) Islet cell purity:79.6%.(2) Islet cell survival rate,compared Group B,Group D with Group A,P＜0.05,α=0.05.statistical difference between Group B,Group D and Group A.compared Group C with Group A,P＞0.05α=0.05.NO statistical difference between them. compared Group C with different time,P＞0.05,in accordance with the standard testα=0.05, no statistical difference between the two.(3) Test Insulin secretion,compared Group B, Group D with Group A,P＜0.05,α=0.05.statistical difference between Group B,Group D and Group A.compared Group C with Group A,P＞0.05α=0.05.NO statistical difference between them.compared Group C with different time,P＞0.05,in accordance with the standard testα=0.05,no statistical difference between the two.Conclusions Vitrification cryopreservation is a useful way that it is used for building islet cells bank.The way is consisted of directly into liquid nitrogen,r apid rewarming at 37℃and appropriate ratio of cryoprotectant.Promote the progress of islet cell transplantation for diabetes patients around the world.