The High Throughput Screening And Testing Kit For Drug Candidate Efficacy And Toxicity Based On C.elegans

Studies have shown that the C.elegans are excellent high throughput screening model.But there are still some problems based on this platform,such as the C.elegans are difficult to culture,high throughput screening methods aren't standardized and so on,which lead to be difficult to use this model in a wild range.The main purpose of this paper is to establish standardized high throughput screening methods and build the corresponding kits.So the development work of three kits is carried out,which can provide some theoretical and technical support for the realization of these kits application in a wide range.The development of high throughput screening kit for discovering new drugs of down-regulating over-activated Ras based on the C.elegans use the wild type ratio as its evaluation indicator,determining the appropriate vehicle is DMSO?2%,PEG-400?2%,Tween-80?3%;determining the appropriate treatment method is drugs treat the MT2124 strain nematods hatching within 48 hours from L1 stage to adult stage;determining the positive control is RBS which the final concentration is 3.2?g/mL;determining the appropriate storage temperature is the time is three days at 4?.So this kit can be assembled as following elements,S medium,MT2124 strain nematods,positive control,NaN₃ and 96 well plate and it can be stored for three days at 4?.The development of high throughput screening kit for discovering new drugs of down-regulating ROS lever based on the C.elegans use ROS lever in vivo as its evaluation indicator,determining the appropriate staining temperature and time of H₂DCF-DA is 20?and30 min respectively;determining the appropriate treatment method is drugs treat the N2 strain nematods hatching within 48 hours from L1 stage to L4 stage;determining the positive control is Vc which the final concentration is 4mM.So this kit can be assembled as following elements,S medium,N2 strain nematods,positive control,H₂DCF-DA and black 384 well plate.Its appropriate vehicle and storage condition also need to be further studied.The development of high throughput testing kit for toxicity of drugs based on the C.elegans use the death rate as its evaluation indicator and mercuric chloride as its treatment drug,determining the appropriate transporter is transparent 384 well plate and there should be about 50nematodes in each well;determining the appropriate treatment method is drugs treat L4 stage N2nematods together with SYTOX Green many C.elegans are dead;determining the appropriate testing method is photographing the fluorescence of dead nematodes.So this kit can be assembled as following elements,K medium,N2 strain nematods,positive control,SYTOX Green and transparent 384 well plate.Its appropriate vehicle and storage condition also need to be further studied.