Scripties UMCG - Rijksuniversiteit Groningen
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Generation of functional single positive T-cells from induced pluripotent stem cells of an ovarian cancer patient

(2018) Sluis, J.M.

Introduction. Ovarian cancer is the number one cause of gynaecological cancer deaths and despite aggressive treatments, the mortality rate is still extremely high. Adoptive T cell therapy is a promising therapy in which T cells are generated in vitro and transferred back to the patient. The European Research institute for Biology of Ageing have established a method to efficiently generate induced pluripotent stem cells from renal epithelial cells found in urine and the usage of these cells would have great advantages, because unlimited patient specific T cells with a large variety of T cell receptors could be developed without the need of invasive treatment. This project aims to generate single positive T cells by differentiating induced pluripotent stem cells derived from an ovarian cancer patient.
Methods. First, induced pluripotent stem cells were differentiated to haematopoietic stem cells via an OP9 co-culture, with cytokines in serum-free medium, GSK3 inhibitors or an combination of these methods. Thereafter, haematopoietic stem cells were differentiated to T cells via an OP9-delta like ligand 1 co-culture or coated delta like ligand 4 protein plates.
Results. OP9 co-culture, GSK3 inhibitors and the combination of these methods generated CD34+CD41a-CD235a- haematopoietic stem cells (0.42-4.02%). When transferred on a delta like ligand 4 coated plate only the haematopoietic stem cells from the combination protocol survived and of these cells 48.6% were differentiated towards CD7+CD5-CD56- cells (immature T cells) and 17.2% towards CD7+CD5-CD56+ cells (natural killer cells). Due to time limits the results of the OP9-delta like ligand co-culture were not measured.
Conclusion. No mature single positive T cells were generated during this project. However, immature DN2 T cells were successfully generated.

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