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Modelling chronic lymphocytic leukaemia (CLL) cell: microenvironment interactions in two- and three-dimensional culture systems

Modelling chronic lymphocytic leukaemia (CLL) cell: microenvironment interactions in two- and three-dimensional culture systems
Modelling chronic lymphocytic leukaemia (CLL) cell: microenvironment interactions in two- and three-dimensional culture systems
Chronic lymphocytic leukaemia (CLL) is a common B-cell malignancy characterised by the Accumulation of malignant B cells in the blood, bone marrow and secondary lymphoid Organs. CLL cells rapidly undergo apoptosis when cultured in vitro, highlighting the Importance of supporting tissue microenvironments for pathogenesis. The survival of CLL Cells depends on a complex interplay of distinct cell types, the extracellular matrix and Soluble factors within the microenvironment and as such modelling the CLL Microenvironment has been an area under intense investigation. This study investigated The use of a novel method, ‘acoustic trapping’ to model the CLL microenvironment. The Overarching hypothesis is that acoustic trapping devices can be used to ‘levitate’ CLL cells Away from artificial surfaces and, with the use of the HFFF2 fibroblast cell line, a three Dimension (3D) model of the CLL:stromal microenvironment can be created to probe Molecular interactions between the two cell types in vitro. The first experiments characterised the use of the HFFF2 cell line for its suitability for use In this project. Results indicated that co-culture with the cell line promoted CLL cell Survival in all samples studied and further experiments demonstrated that CLL cell Survival occurred in a contact independent manner. Optimisation of ‘acoustic trapping Devices’ designed and manufactured by Dr Peter Glynne-Jones (Faculty of Engineering And the Environment, University of Southampton), demonstrated that both CLL and HFFF2 cells could be co levitated to form agglomerates that contracted over a 48 hour Period to form 3D structures. However further investigations revealed that there was a Significant loss of viability of both cell types in the devices and this led to the conclusion That at present, acoustic trapping devices have not been optimised enough to allow for Modelling of the CLL microenvironment. Further experiments concentrated on the characterisation of the HFFF2 co-culture model Using gene expression profiling (GEP) to determine CLL:fibroblast interactions in vitro. Results indicating that there was a distinct gene expression profile in CLL cells that had Been co-cultured with HFFF2 cells or HFFF2-derived conditioned media (CM) resulting in The enrichment of a number of different cytokines and chemokines. Analysis of four Specific cytokines/chemokines revealed that culture with HFFF2 CM resulted in the Upregulation of IL-6, IL-8, CXCL2 and CCL2. Purification of CLL cells lead to the discovery That some were directly induced following culture with HFFF2-CM while the induction of Others required a third intermediate cell type. Overall results demonstrate that the HFFF2 Cell line is a suitable model for investigating CLL/microenvironment interactions and in this Model HFFF2 cells, accessory cells and soluble factors are working together to provide Pro-survival signals to the CLL cells.
University of Southampton
Lemm, Elizabeth Anne
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Lemm, Elizabeth Anne
f363493b-47ce-4f8e-9e94-272486397cb3
Hill, Martyn
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Packham, Graham
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Tare, Rahul
587c9db4-e409-4e7c-a02a-677547ab724a
Thomas, Gareth
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Lemm, Elizabeth Anne (2016) Modelling chronic lymphocytic leukaemia (CLL) cell: microenvironment interactions in two- and three-dimensional culture systems. University of Southampton, Doctoral Thesis, 438pp.

Record type: Thesis (Doctoral)

Abstract

Chronic lymphocytic leukaemia (CLL) is a common B-cell malignancy characterised by the Accumulation of malignant B cells in the blood, bone marrow and secondary lymphoid Organs. CLL cells rapidly undergo apoptosis when cultured in vitro, highlighting the Importance of supporting tissue microenvironments for pathogenesis. The survival of CLL Cells depends on a complex interplay of distinct cell types, the extracellular matrix and Soluble factors within the microenvironment and as such modelling the CLL Microenvironment has been an area under intense investigation. This study investigated The use of a novel method, ‘acoustic trapping’ to model the CLL microenvironment. The Overarching hypothesis is that acoustic trapping devices can be used to ‘levitate’ CLL cells Away from artificial surfaces and, with the use of the HFFF2 fibroblast cell line, a three Dimension (3D) model of the CLL:stromal microenvironment can be created to probe Molecular interactions between the two cell types in vitro. The first experiments characterised the use of the HFFF2 cell line for its suitability for use In this project. Results indicated that co-culture with the cell line promoted CLL cell Survival in all samples studied and further experiments demonstrated that CLL cell Survival occurred in a contact independent manner. Optimisation of ‘acoustic trapping Devices’ designed and manufactured by Dr Peter Glynne-Jones (Faculty of Engineering And the Environment, University of Southampton), demonstrated that both CLL and HFFF2 cells could be co levitated to form agglomerates that contracted over a 48 hour Period to form 3D structures. However further investigations revealed that there was a Significant loss of viability of both cell types in the devices and this led to the conclusion That at present, acoustic trapping devices have not been optimised enough to allow for Modelling of the CLL microenvironment. Further experiments concentrated on the characterisation of the HFFF2 co-culture model Using gene expression profiling (GEP) to determine CLL:fibroblast interactions in vitro. Results indicating that there was a distinct gene expression profile in CLL cells that had Been co-cultured with HFFF2 cells or HFFF2-derived conditioned media (CM) resulting in The enrichment of a number of different cytokines and chemokines. Analysis of four Specific cytokines/chemokines revealed that culture with HFFF2 CM resulted in the Upregulation of IL-6, IL-8, CXCL2 and CCL2. Purification of CLL cells lead to the discovery That some were directly induced following culture with HFFF2-CM while the induction of Others required a third intermediate cell type. Overall results demonstrate that the HFFF2 Cell line is a suitable model for investigating CLL/microenvironment interactions and in this Model HFFF2 cells, accessory cells and soluble factors are working together to provide Pro-survival signals to the CLL cells.

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Published date: July 2016
Organisations: University of Southampton, Cancer Sciences

Identifiers

Local EPrints ID: 410613
URI: http://eprints.soton.ac.uk/id/eprint/410613
PURE UUID: 4dfa8b2a-f71a-4964-9e4c-7fd27210878f
ORCID for Martyn Hill: ORCID iD orcid.org/0000-0001-6448-9448
ORCID for Graham Packham: ORCID iD orcid.org/0000-0002-9232-5691
ORCID for Rahul Tare: ORCID iD orcid.org/0000-0001-8274-8837

Catalogue record

Date deposited: 09 Jun 2017 09:13
Last modified: 14 Mar 2019 01:54

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Contributors

Author: Elizabeth Anne Lemm
Thesis advisor: Martyn Hill ORCID iD
Thesis advisor: Graham Packham ORCID iD
Thesis advisor: Rahul Tare ORCID iD
Thesis advisor: Gareth Thomas

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