Preparation of Cell Model Systems for Digital Spatial Profiling
(2022) KIMM01 20221Department of Immunotechnology
Educational programmes, LTH
- Abstract
- Current cancer treatments can be non-specific, aggressive, and often lead to poor quality of life due to significant side effects. There is a dire need for personalised medicine and accurate tumour subtyping, particularly with cancers with high levels of heterogeneity. Spatial omics is transforming the way in which we understand and treat cancer, paving the way for more patient-specific treatments. The GeoMx Digital Spatial Profiler (DSP) by Nanostring allows for high-plex, reproducible, and non-destructive spatial analysis of protein and RNA by the use of oligo-tags. It provides a detailed analysis of tumour heterogeneity, identifies biomarkers, and monitors tumour progression from patient-derived samples. The GeoMx technology was... (More)
- Current cancer treatments can be non-specific, aggressive, and often lead to poor quality of life due to significant side effects. There is a dire need for personalised medicine and accurate tumour subtyping, particularly with cancers with high levels of heterogeneity. Spatial omics is transforming the way in which we understand and treat cancer, paving the way for more patient-specific treatments. The GeoMx Digital Spatial Profiler (DSP) by Nanostring allows for high-plex, reproducible, and non-destructive spatial analysis of protein and RNA by the use of oligo-tags. It provides a detailed analysis of tumour heterogeneity, identifies biomarkers, and monitors tumour progression from patient-derived samples. The GeoMx technology was developed for formalin-fixed paraffin-embedded (FFPE) tissue or fresh frozen samples but has not previously been used to assess 3D cell models until this study. In this report, a protocol was developed and analysed using three 3D matrices: Hydrogel HyStem®-C, Cellevate cellulose nanofiber, and Geltrex. Co-cultures of lung carcinoma cells (A549) and T lymphoblast cells (Molt-4) were embedded in these matrices. Furthermore, co-culture and A549 monoculture spheroids were generated following 4-day incubation in non-adherent ultra-low absorbance plates. The findings open possibilities for further optimisation and the analysis of more complex 3D cell models, like organoids, on the GeoMx DSP platform paving the way for reliable in-vitro model systems. (Less)
- Popular Abstract
- Cancer is a detrimental disease that affects the lives of millions. There is a constant need for the development of new cancer treatments as a way to prolong the lives of those affected. Unfortunately, the development of drugs is time-consuming, costly, and, more often than not, unsuccessful following pre-screening tests. The main pre-screening steps in getting a drug to market include laboratory testing, animal testing, and clinical trials in humans. The better the pre-screening testing in the laboratory phase, the less time and money spent investigating a drug that might inevitably fail. Furthermore, better laboratory studies will reduce the need for animal testing, which is of primary ethical concern. The focus of this project was to... (More)
- Cancer is a detrimental disease that affects the lives of millions. There is a constant need for the development of new cancer treatments as a way to prolong the lives of those affected. Unfortunately, the development of drugs is time-consuming, costly, and, more often than not, unsuccessful following pre-screening tests. The main pre-screening steps in getting a drug to market include laboratory testing, animal testing, and clinical trials in humans. The better the pre-screening testing in the laboratory phase, the less time and money spent investigating a drug that might inevitably fail. Furthermore, better laboratory studies will reduce the need for animal testing, which is of primary ethical concern. The focus of this project was to create a more reproducible and cheaper test on an instrument, the GeoMx Digital Spatial Profiler, to help better study and screen drugs for cancer.
The instrument that the test will be made for previously only used pieces of tissue taken directly from cancer patients' tumours. These samples are naturally scarce due to the procedure involved with retrieval, and, in turn, screening of drugs using these samples are not possible due to that the tissues have been fixated and thus cells are in a non-viable state. Therefore, cell model systems designed to mimic tumors are more appropriate for drug screening. Cell-based tests can be 2D or 3D, with 2D more commonly used. 2D cell models are merely the cells being grown on a flat surface, therefore unlike the environment in the body. 3D models involve creating an environment similar to that in the body either by putting these cells in a structure similar to the body, known as 3D matrices or by making the cells form clumps, known as spheroids, to create a small tumour-like mass. Both of these methods are developed in this project.
Although there are a lot of 3D matrices in which the cells can be placed to mimic the tumour environment, not all of them might be suitable for the GeoMx. A lab protocol was made to get the samples to a form in which they can be used on the instrument; thin sections of paraffin-embedded samples. In this study, three of these 3D matrices were studied and compared based on how well they worked with the protocol. The matrix known as Hydrogel HyStem®-C was the most favourable and was used to confirm the overall objectives that cell models can be analysed on the GeoMx instrument. Furthermore, the clumps of cell masses, spheroids, were also successfully formed after allowing for growth for 4-days. Future studies would have to be conducted to determine how these spheroids could be used on the GeoMx instrument. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9096071
- author
- Coffey, Christine LU
- supervisor
- organization
- course
- KIMM01 20221
- year
- 2022
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9096071
- date added to LUP
- 2022-07-19 10:09:38
- date last changed
- 2022-07-19 10:09:38
@misc{9096071,
abstract = {{Current cancer treatments can be non-specific, aggressive, and often lead to poor quality of life due to significant side effects. There is a dire need for personalised medicine and accurate tumour subtyping, particularly with cancers with high levels of heterogeneity. Spatial omics is transforming the way in which we understand and treat cancer, paving the way for more patient-specific treatments. The GeoMx Digital Spatial Profiler (DSP) by Nanostring allows for high-plex, reproducible, and non-destructive spatial analysis of protein and RNA by the use of oligo-tags. It provides a detailed analysis of tumour heterogeneity, identifies biomarkers, and monitors tumour progression from patient-derived samples. The GeoMx technology was developed for formalin-fixed paraffin-embedded (FFPE) tissue or fresh frozen samples but has not previously been used to assess 3D cell models until this study. In this report, a protocol was developed and analysed using three 3D matrices: Hydrogel HyStem®-C, Cellevate cellulose nanofiber, and Geltrex. Co-cultures of lung carcinoma cells (A549) and T lymphoblast cells (Molt-4) were embedded in these matrices. Furthermore, co-culture and A549 monoculture spheroids were generated following 4-day incubation in non-adherent ultra-low absorbance plates. The findings open possibilities for further optimisation and the analysis of more complex 3D cell models, like organoids, on the GeoMx DSP platform paving the way for reliable in-vitro model systems.}},
author = {{Coffey, Christine}},
language = {{eng}},
note = {{Student Paper}},
title = {{Preparation of Cell Model Systems for Digital Spatial Profiling}},
year = {{2022}},
}