23 Jul Preparation method of model used for in vitro glioma stem cell drug screening
Abstract
The invention belongs to the technical field of genetic engineering, and more specifically relates to a preparation method of a model used for in vitro glioma stem cell drug screening. The preparation method comprises steps of glioma stem cell separation, glioma stem cell suspension culturing, and glioma stem cell adhesion culturing. The step of glioma stem cell separation comprises following specific steps: 1, glioma tissue is acquired; 2, the glioma tissue is collected, and is subjected to rinsing; 3, the glioma tissue is cut into pieces, digestive enzyme is added, digestion is carried out for 30min at 37 DEG C, and dissociating is carried out every 5min; 4, reaction is stopped, and centrifugation is carried out; 5, washing is carried out; 6, counting, inoculating, and culturing are carried out. The step of glioma stem cell adhesion culturing and freeze preservation comprises following specific steps: 1, after aggregation of cells obtained in the former step, digestion is carried out so as to obtain single cells, and centrifugation is carried out; 2, counting and subcultring are carried out, culturing solution is changed in the next day of subcultring, and then is changed every other day; 3, when density is larger than 90%, digestion into single cells is carried out, and subcultring or freeze preservation is carried out. The primary culturing method of glioma cells is simple, and is convenient in operation, and adherent primary culturing success rate of glioma stem cells is high.
Year: 2017
Country: CN
Doc No: 106609265