title: THE EFFECTS OF A SIMPLE METHOD FOR CRYOPRESERVATION AND THAWING PROCEDURES ON CORD BLOOD DERIVED DC-BASED ESOPHAGEAL CARCINOMA VACCINE.
abstract: BACKGROUND
  Producing sufficient numbers of DCs at one time point and subsequently cryopreserving the generated DCs in ready-for-use aliquots for clinical application is useful in cancer treatment.
  
  
  OBJECTIVE
  To study the effects of a simplified cryopreservation method and thawing procedures acting on the biological characteristics and specific cytotoxic activity of cord blood derived DC-based esophageal carcinoma vaccine.
  
  
  METHODS
  CD34+ hematopoietic stem cells were isolated from cord blood using CD34+ Progenitor Cell Isolation Kit by magnetic cell sorting system (MACS). The CD34+ cells were expanded with cytokines as DCs, and fused with EC109 cells by PEG-3600. The fused cells were transferred to a freezing tube without rate-controlled freezing and stored at -80 degree C for three weeks. During cryopreservation, 2.5% DMSO, 2.5% glucose and 10% FCS at final concentration was used as stock solution. After thawing, cells were assayed for Typan blue viability, morphology, immunophenotypes and T-cell stimulatory capacity, and specific CTL activity.
  
  
  RESULTS
  Cryopreservation does not cause significant changes in the phenotypes expression or morphology of the fused cells, and the viability were well preserved (Typan blue viability was 77.2±1.8%). After being stimulated by DC-based esophageal carcinoma vaccine either before or after cryopreservation, the numbers of CD3+T/CD4+T and CD3+T/CD8+T lymphocytes increased obviously, especially for CD3+T/CD4+T, and the ratio of CD4/CD8 changed from 0.85 to 1.29 and 1.25 respectively. Specific CTL activity were well preserved (compare to the fresh fused vaccine, P>0.05).
  
  
  CONCLUSIONS
  A simple -80 degree C freezing and storage method is practical for cord blood derived DC-based esophageal carcinoma vaccine. It will greatly facilitate the clinical use of DC-based vaccine for immunotherapy.
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.
authors.forename: J
authors.initials: J
authors.lastname: Yu
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.
authors.forename: L
authors.initials: L
authors.lastname: Xie
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.
authors.forename: S
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Shantou University Medical College, Shantou, China.
authors.forename: G
authors.initials: G
authors.lastname: Guo
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Shantou University Medical College, Shantou, China. bingmingchen@sina.com.
authors.forename: B
authors.initials: B
authors.lastname: Chen
mesh_terms: D019496:Cancer Vaccines / Q000276:immunology*; D002277:Carcinoma / Q000276:immunology*; D015925:Cryopreservation / Q000379:methods*; D003601:Cytotoxicity Tests, Immunologic; D003713:Dendritic Cells / Q000502:physiology*; D004938:Esophageal Neoplasms / Q000276:immunology*; D006801:Humans
chemical_list: D019496:Cancer Vaccines