[The induction and function of dendritic cells from human cord blood with IFN-alpha and GM-CSF in vitro].
OBJECTIVE
To probe into the possibility of obtaining dendritic cells (DCs) from human cord blood with rhIFN-alpha and rhGM-CSF.


METHODS
The plastic adherent cells (monocyte-rich cells) from cord blood were cultured with rhGM-CSF and rhIFN-alpha for 7 days. The morphological properties of DCs were observed by microscopy, the expression of CD83 and CD86 on DCs were determined by flow cytometry. The function of DCs stimulating allogeneic T lymphocyte proliferation was detected by MTT colorimetric method.


RESULTS
After 7 days culture, some of the cultured cells acquired typical DC morphology and showed increased expression of CD83 and CD86. The yield of DCs attained its peak when the concentrations of rhIFN-alpha and rhGM-CSF were 600 U/ml and 2000 U/ml, respectively. When cultured cells (containing DCs) as stimulator cells were co-cultured with allogeneic T lymphocytes (effector cells) in different ratio, they could stimulate T cell proliferation remarkably, especially when the ratio of effector cells to stimulator cells was 20 to 1.


CONCLUSIONS
Our findings suggest that cord blood adherent cells, when cultured with rhGM-CSF and rhIFN-alpha, can be induced into mature DCs with the function of stimulating allogeneic T cell proliferation.
Department of Paediatrics, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Dong-ling
DL
Hong
Qiang
Q
Li
Cang-song
CS
Jia
D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology / Q000502:physiology; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology; D013601:T-Lymphocytes / Q000166:cytology
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor