title: [Comparison of Ex Vivo Expanded and Highly Purified NK Cell-Mediated Cytotoxicity Detected by 3 Different Staining Methods of Flow Cytometry].
abstract: OBJECTIVE
  To compare the cytotoxicity of ex vivo expanded NK cells detected by flow cytometry with 3 different staining methods.
  
  
  METHODS
  NK cells were collected from peripheral blood on the 17thday after culture. The cultured cells were divided into 3 groups: group A , B, and C. The cells in group A were stained with CFSE/Annectin-V/7-AAD; the cells in group B were stained with Annectin-V/PI, and the cells in group C cells were stained with CFSE/PI. The E:T ratios in 3 groups were 10:1, 20:1 and 40:1, respectively, the K562 cells were incubated with NK cells for 4 hrs.
  
  
  RESULTS
  The purity of NK cells(CD3-CD56+) reached to (16.34±10.51)% on day 0 and to (83.63±10.63)% on the day 17 after incubation(P<0.05); the cytotoxicity of group A was significantly higher than thay of group B at different E:T ratio (P<0.05). The cytotoxicities in A, B, C groups at E:T ratio=10:1 were (36.56±3.69)%, (10.85±2.09)% and (22.35±2.71)% respectively; the cytotoxicities in A, B, C groups at E:T ratio=9:1 were (47.83±5.52)%, (39.07±5.55)% and (29.61±4.81)%; the cytotoxicities in A, B, C groups at E:T ratio=40:1 were (67.7±4.77)%, (51.51±4.43)% and (44.12±5.62)% respectively. Meanwhile, the cytotoxicity in group A was significantly higher than that in group C at different E:T ratio (P<0.05), the percentage of cytotoxicity was (36.56±3.69)% vs (10.85±2.09)%, (47.83±5.52)% vs (29.61±4.81)%, (67.7±4.77)% vs (44.12±5.62)%, respectively.
  
  
  CONCLUSIONS
  CFSE/Annectin-V/7-AAD is able to clearly show human NK cell cytotoxicity against human tumors. Moreover, this staining technique also allows to distinguish different stages of cytotoxic killing as early and late apoptotic phase.
authors.affiliation: Department of Biotherapy, Shanxi Dayiyuan, Shanxi Academy of Medical Sciences,Taiyuan 030032, Shanxi Province, China.
authors.forename: Ji-Qiang
authors.initials: JQ
authors.lastname: Guo
authors.affiliation: Department of Biotherapy, Shanxi Dayiyuan, Shanxi Academy of Medical Sciences,Taiyuan 030032, Shanxi Province, China.
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Han
authors.affiliation: Department of Biotherapy, Shanxi Dayiyuan, Shanxi Academy of Medical Sciences,Taiyuan 030032, Shanxi Province, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Biotherapy, Shanxi Dayiyuan, Shanxi Academy of Medical Sciences,Taiyuan 030032, Shanxi Province, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: Department of Biotherapy, Shanxi Dayiyuan, Shanxi Academy of Medical Sciences,Taiyuan 030032, Shanxi Province, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Jia
authors.affiliation: Postgraduat School, Shanxi Medical University, Taiyuan 030032, Shanxi Province, China.
authors.forename: Bao-Li
authors.initials: BL
authors.lastname: Jin
authors.affiliation: Department of Biotherapy, Shanxi Dayiyuan, Shanxi Academy of Medical Sciences,Taiyuan 030032, Shanxi Province, China. E-mail:13994204099@163.com.
authors.forename: Jun-Ping
authors.initials: JP
authors.lastname: Zhang
mesh_terms: D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry; D006801:Humans; D020014:K562 Cells; D007694:Killer Cells, Natural; D013194:Staining and Labeling