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Amplification of T cells from human cord blood in serum-deprived culture stimulated with stem cell factor, interleukin-7 and interleukin-2

Bone Marrow Transplantation volume 31, pages 713–723 (2003)Cite this article

Summary:

We report the effects exerted by cytokine combinations, including stem cell factor (SCF), interleukin-7, interleukin-4 and interleukin-2, on the amplification of T cells from cord blood (CB) mononuclear cells cultured for 10–11 days under serum-deprived conditions. Of all the combinations investigated, SCF+interleukin-7 sustained the best fold increase (FI) of total nucleated cells (FI=6.4±1.17), amplifying preferentially CD4+ over CD8+ T-cell subsets (FI=4.72±0.79 vs 2.73±1.2, respectively, P<0.05). The addition of interleukin-2 to this combination did not significantly increase the total number of cells generated (FI=7.4±2.27), but allowed preferential amplification of CD8+ over CD4+ T cells (FI=6.04±0.14 vs 1.67±0.6, respectively, P<0.05). Single-strand conformation polymorphism analysis of the T-cell receptor Vβ-chain rearrangements expressed by the expanded T cells indicated that the complexity of the T-cell repertoire had increased after 10 days of culture in the presence of SCF and IL-7. Interestingly, a modest expansion (FI=8.67±1.5) of myeloid progenitor cells was also observed in these cultures. These results indicate that it is possible to expand specific T-cell subsets for adoptive immunotherapy without losing myeloid progenitor cells necessary for neutrophil recovery after CB transplantation, by modulating the cytokines added to the cultures.

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Acknowledgements

We thank Dr M Marceca (II Clinica Ostetrica, Università agli Studi ‘La Sapienza’, Rome, Italy) for providing CB samples. This work was supported by institutional funds of Istituto Superiore di Sanita', Progetto Finalizzato 1%, Ricerca Corrente and Progetti di Ricerca di Interesse Nazionale 2000, from the Ministry of Health; Progetto Strategico Oncologia CNR-MIUR legge 449/99 and Grant no. E1172 from the Telethon Foundation.

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Authors and Affiliations

  1. Laboratory of Cell Biology, Istituto Superiore di Sanita', Rome, Italy

    M Sanchez, E Alfani & G Migliaccio

  2. Clinical Biochemistry, Istituto Superiore di Sanita', Rome, Italy

    A R Migliaccio

  3. Universita' ‘G. D’Annunzio', Chieti, Italy

    A R Migliaccio & G Migliaccio

  4. Ospedale Spirito Santo, Pescara, Italy

    T Bonfini

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Sanchez, M., Alfani, E., Migliaccio, A. et al. Amplification of T cells from human cord blood in serum-deprived culture stimulated with stem cell factor, interleukin-7 and interleukin-2. Bone Marrow Transplant 31, 713–723 (2003). https://doi.org/10.1038/sj.bmt.1703904

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