title: [Application of vital dye CFDA-SE and SNARF-1 to evaluate mixed lymphocyte reaction].
abstract: OBJECTIVE
  To establish a new method for more comprehensive evaluation of responsive intensity in one-way mixed lymphocyte reaction (MLR).
  
  
  METHODS
  CFDA-SE labeled lymphocytes from lymph nodes of BALB/cJ mice were applied as responder cells, and SNARF-1 labeled splenocytes from BALB/cJ or C57BL/6 mice which had been pretreated with mitomycin C were applied as stimulator cells to establish mixed lymphocyte culture (MLC). After 96 h of culture, cells were harvested and analyzed by flow cytometry to acquire the proliferative information of SNARF-1 negative and CFSE positive staining responder cells. ModFit software was then used to get more proliferation related index.
  
  
  RESULTS
  With increasing division cycles, the CFSE fluorescence was diluted to the extent that the responder cells could not be discriminated from stimulator cells, which was resolved by gating SNARF-1 positive cells out to define responder cells. Several important proliferation related indexes were obtained by ModFit software, such as precursor frequency and proliferation index, etc.
  
  
  CONCLUSIONS
  Our results show that application of vital dye CFDA-SE and SNARF-1, combined with flow cytometry is a powerful tool for evaluation of responsive intensity in one-way MLR.
authors.affiliation: Key Laboratory of Ministry of Education for Tissue Transplantation and Immunology, Jinan University, Guangzhou 510630, China. zhaojingxian@126.com
authors.forename: Jing-xian
authors.initials: JX
authors.lastname: Zhao
authors.forename: Yao-ying
authors.initials: YY
authors.lastname: Zeng
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Xian-hui
authors.initials: XH
authors.lastname: He
mesh_terms: D000818:Animals; D001578:Benzopyrans; D005434:Flow Cytometry; D005452:Fluoresceins / Q000494:pharmacology; D005456:Fluorescent Dyes; D008213:Lymphocyte Activation / Q000187:drug effects; D007959:Lymphocyte Culture Test, Mixed; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009284:Naphthols; D012235:Rhodamines; D013194:Staining and Labeling / Q000379:methods; D013388:Succinimides / Q000494:pharmacology
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D001578:Benzopyrans; D005452:Fluoresceins; D005456:Fluorescent Dyes; D009284:Naphthols; D012235:Rhodamines; D013388:Succinimides; C091666:seminaphthorhodaminefluoride