Culture Characters And Cryopreservation Method Of Caenorhabditis Elegans

C.elegans is very commonly used a parasite in a laboratory, because before this laboratory will c.elegans in the 30% glycerin in liquid nitrogen,-80℃the preservation and in the 10% of glycerol in liquid nitrogen cryopreservation is not successful, in order to using for a long time, has been to subculture, is very trouble. And, recently, some countries have begun to use of c.elegans to study the mechanism of programmed cell death,to develop new ways to treat diseases such as malignant tumor,also will be on the way to treat cancer and AIDS play an important role.In addition by the c.elegans slow aging and prolong life mechanism,is expected to be applied to other animals,based on the results of the study on the treatment of human disease and prolong the life of human itself play a key role.So,in this paper,the author studies on c.elegans the after cryopreservation method is for the purpose of the study of c.elegans plays a fundamental role in various fields,for further in-depth study for the test of c.elegans.This experiment USES the glycerin(Gly),dimethyl sulfoxide(DMSO),ethylene glycol(EG),Glucose(Glucose),albumin five freeze protection liquid comparison test,freezing temperature- 20℃ and-80℃,respectively,and the L1 and L2 stage larvae,larvae stage Dauer larvae and L3 and L4 stage larva and adult cryopreserved comparison.This experiment adopts OP50 e.coli,using the NGM culture medium for culture.The test results:Frozen a month, glycerol,dimethyl sulfoxide, ethylene glycol as cryoprotectant at-80℃ condition, containing L1,L2 Dauer larvae and larvae of worms and larvae contain L1,L2 larvae,L3 larvae,L4 larvae and adult worms can survive a lot;Three months frozen,containing L1,L2 larvae and Dauer larvae cryopreservation survival than larvae containing L1,L2 larvae,L3 larvae,L4 larvae and adult survival rates higher cryopreservation,containing 10% glycerol in PBS containing 10% dimethyl sulfoxide in PBS at-80℃ under the conditions of survival are parasites up to 70%;Six frozen,containing L1,L2 larvae and Dauer larvae cryopreservation survival than larvae containing L1,L2 higher larvae,L3 larvae,L4 larvae and adults cryopreservation survival the use of dimethyl sulfoxide containing 10% of PBS was frozen in effect under the conditions of-80 ℃ best,containing L1, L2 larvae and Dauer larvae cryopreservation survival rate of 65%.the at-80℃,use PBS solution containing 10% dimethyl sulfoxide cryopreserved effect is best,L1 and L2 stage larvae and Dauer larvae cryopreservation of survival rate is higher.Hence,The test on c.elegans is very important for further in-depth study.