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In vitro differentiation of natural killer T cells from human cord blood CD34+ cells

Cited 11 time in Web of Science Cited 10 time in Scopus
Issue Date
2003-04
Citation
British Journal of Haematology, Vol.121 No.1, pp.148-156
Keywords
NKT cellscord bloodCD34(+) cellsinterleukin 15differentiation
Abstract
Natural killer T (NKT) cells are involved in innate immune defence and also in the regulation of adaptive immune responses. However, the development of NKT cells in vitro has not been fully characterized and culture conditions have not been fully optimized. In the present study, we found that an NKT cell fraction developed during the in vitro culture of cord blood (CB) CD34(+) cells, and this was subsequently characterized both phenotypically and morphologically. CD34(+) cells purified from 10 human CB were cultured in the presence of several cytokines and analysed by flow cytometry, light microscopy and electron microscopy. The NKT cell fraction, defined phenotypically (CD3(+) CD16(+) CD56(+) CD94(+) ) as expressing the invariant T-cell receptor Valpha24 and Vbeta11, appeared in the CD56(hi) fractions. Intracytoplasmic staining demonstrated that interferon-gamma and interleukin 4 (IL-4) were detected in the CD56(hi) fractions. IL-15 was essential and, in combination with either flt3-ligand (FL) or stem cell factor (SCF), was sufficient to induce the development of NKT cells. The phenotype of the NKT cell fraction was CD45RO(+) CD45RA(-) and CD4(+) CD8alpha(+) . Morphologically, they were very large, with either round or oval nuclei, moderately condensed chromatins, voluminous weakly basophilic cytoplasm and various cytoplasmic granules such as dense core granules, multivesicular bodies, and intermediate form granules. When CD34(+) cells purified from bone marrow (BM) were compared with those from CB, the latter were consistently more efficient at generating CD56(hi) NKT cell fractions. In conclusion, IL-15 in combination with FL and/or SCF can induce the differentiation of NKT cells from human CB CD34(+) cells.
ISSN
0007-1048
URI
https://hdl.handle.net/10371/177159
DOI
https://doi.org/10.1046/j.1365-2141.2003.04230.x
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College of Medicine/School of Medicine (의과대학/대학원)Internal Medicine (내과학전공)Journal Papers (저널논문_내과학전공)
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