The　drug　development　process　involves　a　variety　of　drug　activity　evaluations,　which　can　determine　drug　efficacy,　strictly　analyze　the　biological　indicators　after　the　drug　action,　and　use　these　indicators　as　the　preclinical　drug　evaluation　criteria.　At　present,　most　of　the　screening　of　preclinical　anticancer　drugs　mainly　relies　on　traditional　2D　cell　culture.　However,　this　traditional　technology　cannot　simulate　the　tumor　microenvironment　in　vivo,　let　alone　reflect　the　characteristics　of　solid　tumors　in　vivo,　and　has　a　relatively　poor　ability　to　predict　drug　activity.　3D　cell　culture　is　a　technology　between　2D　cell　culture　and　animal　experiments,　which　can　better　reflect　the　biological　state　in　vivo　and　reduce　the　consumption　of　animal　experiments.　3D　cell　culture　can　link　the　individual　study　of　cells　with　the　study　of　the　whole　organism,　reproduce　in　vitro　the　biological　phenotype　of　cells　in　vivo　more　greatly,　and　thus　predict　the　activity　and　resistance　of　anti-tumor　drugs　more　accurately.　In　this　paper,　the　common　techniques　of　3D　cell　culture　are　discussed,　with　emphasis　on　its　main　advantages　and　application　in　the　evaluation　of　anti-tumor　resistance,　which　can　provide　strategies　for　the　screening　of　anti-tumor　drugs.