title: [Change Trend of Viablity, Apoptosis and ROS Production of Neutrophil in vitro Culture].
abstract: OBJECTIVE
  To explore the change trend of cell viability, apoptosis and reactive oxygen species (ROS) production of peripheral blood neutrophils in in vitro culture.
  
  
  METHODS
  Neutrophils were isolated by density gradient centrifugation and cultured in medium containing 10% fetal bovine serum for diffentrent times in a fall humidified atmosphere with 5% CO2at 37 °C. Cell viability, spontaneous apoptosis and ROS level were detected by flow cytometry at 0, 24, 48, 72 and 96 hours.
  
  
  RESULTS
  The cell viability of neutrophils could be maintained above 90% at 24 h in culture in vitro, and there was no significant difference between culture in vitro at 0 h and 24 h (P>0.05), but after culture in vitro at 72 h, the cell viability of neutrophils obviously droped, the count of apoptotic cells increased, and after culture in vitro at 96 h the neutrophils almostly all died and the production of ROS at 12 h significantly decreased as compared with that at 0 h.
  
  
  CONCLUSIONS
  The cell viability of neutrophils at 24 h cultures in vitro can be maintained relative stable with cell viability above 90% and unobvious change of count of apoptotic cells, but the neutrophil function test of ROS should be carried out immediately after cell separation.
authors: Central Laborontory of Lianyungang Hospital Affiliated to Xuzhou Medical College. Lianyungang 22200, Jiangsu Province, China; Teaching and Research Section of Clinical Diagnosis, Kangda School of Nanjing Medlical University, Lianyungang 222000, Jiangsu Province, China.
authors: Hai-Ning
authors: HN
authors: Li
authors: Central Laborontory of Lianyungang Hospital Affiliated to Xuzhou Medical College. Lianyungang 22200, Jiangsu Province, China.
authors: Shao-Lin
authors: SL
authors: Zhao
authors: Central Laborontory of Lianyungang Hospital Affiliated to Xuzhou Medical College. Lianyungang 22200, Jiangsu Province, China.
authors: Jin
authors: J
authors: Yang
authors: Central Laborontory of Lianyungang Hospital Affiliated to Xuzhou Medical College. Lianyungang 22200, Jiangsu Province, China. E-mail: medical1121@126.com.
authors: Wei
authors: W
authors: Li
mesh_terms: D000818:Animals; D017209:Apoptosis; D002417:Cattle; D002469:Cell Separation; D002470:Cell Survival; D005434:Flow Cytometry; D009504:Neutrophils; D017382:Reactive Oxygen Species
chemical_list: D017382:Reactive Oxygen Species