Freeze-thawing procedures have no influence on the phenotypic and functional development of dendritic cells generated from peripheral blood CD14+ monocytes.
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.
Hori
Shin-ichi
S
Hematopoietic Stem Cell Transplant/Immunotherapy Unit, National Cancer Center Hospital, Tokyo, Japan.
Heike
Yuji
Y
Takei
Masao
M
Maruyama
Midori
M
Inoue
Yoshiko
Y
Lee
Je-Jung
JJ
Kim
Hyeoung-Joon
HJ
Harada
Yukie
Y
Kawai
Hiroyuki
H
Shimosaka
Akihiro
A
Kami
Masahiro
M
Tanosaki
M D Ryuji
MD
Wakasugi
Hiro
H
Saito
Shigeru
S
Takaue
Yoichi
Y
Kakizoe
Tadao
T
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
D003470:Culture Media; D003911:Dextrans; D018950:Lipopolysaccharide Receptors; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor