title: Freeze-thawing procedures have no influence on the phenotypic and functional development of dendritic cells generated from peripheral blood CD14+ monocytes.
abstract: Little is known about the potential influence of cryopreservation on the biologic activities of dendritic cells (DCs). In this study, we examined the effects of freeze-thawing on the phenotypic and functional development of human DCs obtained from granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood CD14+ cells. CD14+ cells were cultured, immediately or after freeze-thawing, with granulocyte-macrophage CSF and interleukin-4 for 9 days, and then with added tumor necrosis factor-alpha for another 3 days. For both fresh and freeze-thawed monocytes, immature DCs harvested on day 6 and mature DCs harvested on day 9 of culture were examined under the same conditions. Cells were compared with regard to their 1) capacities for antigen endocytosis and chemotactic migration (immature DCs), and 2) allogeneic mixed lymphocyte reaction and antigen-specific cytotoxic T lymphocyte responses (mature DCs). Freeze-thawing did not affect the viability or subsequent maturation of DCs at any stage of development. Furthermore, essentially no difference was observed in phenotype or function between cells generated from fresh or cryopreserved/thawed cells. Although this study design was limited with the use of fetal bovine serum, the observation still suggests that freeze-thawing does not affect viability, phenotype, subsequent maturation, or functions of DCs at any stage of maturation.
authors.affiliation: Hematopoietic Stem Cell Transplant/Immunotherapy Unit, National Cancer Center Hospital, Tokyo, Japan.
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Hori
authors.forename: Yuji
authors.initials: Y
authors.lastname: Heike
authors.forename: Masao
authors.initials: M
authors.lastname: Takei
authors.forename: Midori
authors.initials: M
authors.lastname: Maruyama
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Inoue
authors.forename: Je-Jung
authors.initials: JJ
authors.lastname: Lee
authors.forename: Hyeoung-Joon
authors.initials: HJ
authors.lastname: Kim
authors.forename: Yukie
authors.initials: Y
authors.lastname: Harada
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kawai
authors.forename: Akihiro
authors.initials: A
authors.lastname: Shimosaka
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kami
authors.forename: M D Ryuji
authors.initials: MD
authors.lastname: Tanosaki
authors.forename: Hiro
authors.initials: H
authors.lastname: Wakasugi
authors.forename: Shigeru
authors.initials: S
authors.lastname: Saito
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Takaue
authors.forename: Tadao
authors.initials: T
authors.lastname: Kakizoe
mesh_terms: D002478:Cells, Cultured; D015925:Cryopreservation; D003470:Culture Media; D003713:Dendritic Cells / Q000276:immunology* / Q000502:physiology; D003911:Dextrans / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000502:physiology; D006801:Humans; D015847:Interleukin-4 / Q000502:physiology; D018950:Lipopolysaccharide Receptors / Q000097:blood*; D008954:Models, Biological; D009000:Monocytes / Q000276:immunology*; D010641:Phenotype; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D003470:Culture Media; D003911:Dextrans; D018950:Lipopolysaccharide Receptors; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor