An optimized disaggregation method for human lung tumors that preserves the phenotype and function of the immune cells

[摘要] Carefulpreparationofhumantissuesisthecornerstoneofobtainingaccuratedatainimmunologicstudies.Despitetheessentialimportanceoftissueprocessingintumorimmunologyandclinicalmedicine,currentmethodsoftissuedisaggregationhavenotbeenrigorouslytestedfordatafidelity.Thus,wecriticallyevaluatedthecurrenttechniquesavailableintheliteraturethatareusedtopreparehumanlungtumorsforimmunologicstudies.WediscoveredthattheseapproachesaresuccessfulatdigestingcellularattachmentsandECMs;however,thesemethodsfrequentlyaltertheimmunecellcompositionand/orexpressionofsurfacemolecules.Wethusdevelopedanovelapproachtopreparehumanlungtumorsforimmunologicstudiesbycombininggentlemechanicalmanipulationwithanoptimizedcocktailofenzymesusedatlowdoses.Thisenzymaticdigestioncocktailoptimizedcellyieldandcellviability,retrievedallmajortumor‐associatedcellpopulations,andmaintainedtheexpressionofcell‐surfacemarkersforlineagedefinitionandinvivoeffectorfunctions.Toourknowledge,wepresentthefirstrigorouslytesteddisaggregationmethoddesignedforhumanlungtumors...

[学科分类] 生理学

[关键词] enzymatic digestion;collagenase;tumor‐infiltrating immune cells