Luevano Salinas, ME;
(2013)
Generation of Natural Killer Cells from Umbilical Cord Blood Stem Cells, Characterisation and Application for Immunotherapy.
Doctoral thesis (PhD), UCL (University College London).
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Abstract
Adoptive (Natural killer) NK cell therapy relies on the use of a large amount of NK cells that are cytotoxic, and yet not exhausted. For this purpose, NK cells can be isolated from cord blood, peripheral blood or generated in vitro from haematopoietic stem cells (HSC). In vitro generation of high numbers of activated NK cells using HSC would facilitate multiple infusions and treatment of patients with large tumour burden, allowing to by-pass the limitations of NK cell numbers and activation state of blood-derived NK cells. However, comprehensive studies about the use of fresh or cryopreserved HSC and of different HSC sources for protocols of NK cell production in vitro have yet to be performed. The aim of this thesis was to investigate these variables and establish an optimal protocol for the generation of NK cells in vitro. This work investigated the use of a published protocol and a modified version using only IL-15 for the last weeks of culture; moreover, the comparison of NK cells derived from fresh cord blood stem cells (CBSC) and frozen CBSC and a different HSC source, mobilised peripheral blood stem cells (PBSC), was performed. Using this protocol, we showed that frozen CBSC generated higher NK cell numbers expressing activating receptors, lacking killer-cell immunoglobulin-like receptor expression but with better immunoregulatory and cytotoxic properties compared to NK cells from fresh CBSC and PBSC cultures. More than half of the NK cells generated in vitro from all HSC types expressed the myeloid-marker CD33; blocking of this marker did not impact on NK cell functions. Finally, CBSC and PBSC showed a different threshold for NK cell activation with interleukin 12. In conclusion, our study provides evidence that frozen CBSC are a suitable source of HSC for NK cell generation in vitro compared to fresh CBSC and frozen PBSC.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | PhD |
| Title: | Generation of Natural Killer Cells from Umbilical Cord Blood Stem Cells, Characterisation and Application for Immunotherapy |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Third party copyright material has been removed from ethesis. |
| Keywords: | Stem cells, Immunotherapy, Natural killer cells |
| UCL classification: | UCL > Provost and Vice Provost Offices UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1411341 |
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