The experimental study on inducing and expanding T/NK cells from mononuclear cells of human umbilical cord blood / 中华血液学杂志
Chinese Journal of Hematology
;
(12): 576-579, 2003.
Artigo
em Chinês
| WPRIM
| ID: wpr-354822
ABSTRACT
<p><b>OBJECTIVE</b>To explore the most efficient culture system which can induce cord blood (CB)-mononuclear cells (MNC) to differentiate into mature T/NK cells in vitro.</p><p><b>METHODS</b>The CB MNCs were cultured in six culture systems respectively for 4 weeks. The T/NK cell surface phenotypes were analyzed by flow cytometry and the absolute numbers of nucleated cells (NCs) were counted at each time point. Moreover, cell morphology was identified by Giemsa-Wright staining, and cytotoxicity of the cultured cells to K562 and Raji tumor cells was also evaluated by MTT method.</p><p><b>RESULTS</b>Cultured in the cytokine cocktail of SCF + FLT-3L + IL-7 + IL-15 + TNF-alpha + IL-2, the NCs were (20 approximately 26) x 10(6)/ml in numbers at day 22. The percentage of lymphocytes in the NCs and that of CD(3)(+) T cells in the lymphocytes both exceeded 90% at the same time. Most of the CD(3)(+) T cells were CD(3)(+)CD(8)(+) and the percentage of CD(3)(+)CD(4)(+) T cells declined gradually. The percentage of CD(3)(+)CD(56)(+) NKT cells and gamma delta(+)T cells in the lymphocytes arised from lower than 2% to 30% approximately 40% and 10% approximately 15%, respectively. CD(3)(-)CD(56)(+) NK cells were not expanded. The cytotoxic activity of the cultured cells to K562 and Raji cells at an effectortarget (ET) ratio of 501 was over 75% and about 32% approximately 65%, respectively.</p><p><b>CONCLUSION</b>The most efficient culture system which can induce CB MNC to differentiate into mature T/NK cells in vitro is the cytokines cocktail of SCF + FLT-3L + IL-7 + IL-15 + TNF-alpha + IL-2, and the optimum culture time is 22 days.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Farmacologia
/
Leucócitos Mononucleares
/
Células Matadoras Naturais
/
Linfócitos T
/
Diferenciação Celular
/
Divisão Celular
/
Interleucina-2
/
Fator de Necrose Tumoral alfa
/
Receptores de Antígenos de Linfócitos T gama-delta
/
Biologia Celular
Tipo de estudo:
Estudo prognóstico
Limite:
Humanos
/
Recém-Nascido
Idioma:
Chinês
Revista:
Chinese Journal of Hematology
Ano de publicação:
2003
Tipo de documento:
Artigo
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