Hypothermic preservation of cellular products for advanced therapies
<p>Cell and gene therapies (CGT) are showing positive results in managing a range of healthcare challenges, and one of the exciting examples is the successful treatment of relapsed acute lymphoblastic leukaemia using adoptive transfer of chimeric antigen receptor (CAR) T cells. Many of the pro...
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| Format: | Thesis |
| Language: | English |
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2019
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| _version_ | 1797098179266609152 |
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| author | Nyland, RW |
| author2 | Cui, Z |
| author_facet | Cui, Z Nyland, RW |
| author_sort | Nyland, RW |
| collection | OXFORD |
| description | <p>Cell and gene therapies (CGT) are showing positive results in managing a range of healthcare challenges, and one of the exciting examples is the successful treatment of relapsed acute lymphoblastic leukaemia using adoptive transfer of chimeric antigen receptor (CAR) T cells. Many of the processes in CGT product preparation and administration require optimisation, with the preservation of the therapeutic products being identified as a main challenge. Cryopreservation is the current practice, but it leads to losses in viability and functionality. </p>
<p>This research focussed on development of a novel process, hypothermic preservation, for short-term preservation (days rather than months) between manufacture and application to the patient at the clinic to avoid cryopreservation during transport. This study adopted Design of Experiments (DoE) as a method to rigorously establish a suitable base preservation medium using a fractional factorial design to minimise the number of trials performed, and assessing the responses by viable cell counts and measurement of metabolic activity at the end of the preservation period. Components with negative effects were identified and discarded, which significantly improved the preservation outcome. Optimisation of the medium composition by central composite DoE methods found that antioxidants played a role in the metabolic response of T cells after hypothermic preservation but were unable to maintain the cell functions themselves. Pre-treating T cells with interventions to inhibit cell proliferation forcibly reduced their metabolic activity before exposure to hypothermia and maintained complete functions after recovery. An investigation into cell sedimentation found that gelatin could avoid cell sedimentation but effectively paused T cell activity, which did not recover after preservation. The Ficoll® PM70 additive only slightly reduced the settling velocity but stimulated T cell activity on recovery. The experimental results indicated, however, that sedimentation during preservation did not negatively affect T cell function. A mass transfer computational model showed that oxygenation by conventional cell culture methods maintained suitable oxygen concentrations during preservation. A final study investigated the limitation on the duration of the proposed preservation methods.</p>
<p>Overall, it was concluded that T cells could be preserved up to 3 days using hypothermic preservation at 4oC if their proliferation is paused before the procedure.
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| first_indexed | 2024-03-07T05:05:54Z |
| format | Thesis |
| id | oxford-uuid:d9e7c09d-8943-402a-9574-90384434d338 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T05:05:54Z |
| publishDate | 2019 |
| record_format | dspace |
| spelling | oxford-uuid:d9e7c09d-8943-402a-9574-90384434d3382022-03-27T08:59:20ZHypothermic preservation of cellular products for advanced therapiesThesishttp://purl.org/coar/resource_type/c_db06uuid:d9e7c09d-8943-402a-9574-90384434d338Cryopreservation of organs, tissues, etcT cellsEnglishHyrax Deposit2019Nyland, RWCui, Z<p>Cell and gene therapies (CGT) are showing positive results in managing a range of healthcare challenges, and one of the exciting examples is the successful treatment of relapsed acute lymphoblastic leukaemia using adoptive transfer of chimeric antigen receptor (CAR) T cells. Many of the processes in CGT product preparation and administration require optimisation, with the preservation of the therapeutic products being identified as a main challenge. Cryopreservation is the current practice, but it leads to losses in viability and functionality. </p> <p>This research focussed on development of a novel process, hypothermic preservation, for short-term preservation (days rather than months) between manufacture and application to the patient at the clinic to avoid cryopreservation during transport. This study adopted Design of Experiments (DoE) as a method to rigorously establish a suitable base preservation medium using a fractional factorial design to minimise the number of trials performed, and assessing the responses by viable cell counts and measurement of metabolic activity at the end of the preservation period. Components with negative effects were identified and discarded, which significantly improved the preservation outcome. Optimisation of the medium composition by central composite DoE methods found that antioxidants played a role in the metabolic response of T cells after hypothermic preservation but were unable to maintain the cell functions themselves. Pre-treating T cells with interventions to inhibit cell proliferation forcibly reduced their metabolic activity before exposure to hypothermia and maintained complete functions after recovery. An investigation into cell sedimentation found that gelatin could avoid cell sedimentation but effectively paused T cell activity, which did not recover after preservation. The Ficoll® PM70 additive only slightly reduced the settling velocity but stimulated T cell activity on recovery. The experimental results indicated, however, that sedimentation during preservation did not negatively affect T cell function. A mass transfer computational model showed that oxygenation by conventional cell culture methods maintained suitable oxygen concentrations during preservation. A final study investigated the limitation on the duration of the proposed preservation methods.</p> <p>Overall, it was concluded that T cells could be preserved up to 3 days using hypothermic preservation at 4oC if their proliferation is paused before the procedure. </p> |
| spellingShingle | Cryopreservation of organs, tissues, etc T cells Nyland, RW Hypothermic preservation of cellular products for advanced therapies |
| title | Hypothermic preservation of cellular products for advanced therapies |
| title_full | Hypothermic preservation of cellular products for advanced therapies |
| title_fullStr | Hypothermic preservation of cellular products for advanced therapies |
| title_full_unstemmed | Hypothermic preservation of cellular products for advanced therapies |
| title_short | Hypothermic preservation of cellular products for advanced therapies |
| title_sort | hypothermic preservation of cellular products for advanced therapies |
| topic | Cryopreservation of organs, tissues, etc T cells |
| work_keys_str_mv | AT nylandrw hypothermicpreservationofcellularproductsforadvancedtherapies |