Abstract
Despite extensive research, little progress has been made in glioblastoma therapy, owing in part to a lack of adequate preclinical in vivo models to study this disease. To mitigate this, primary patient-derived cell lines, which maintain their specific stem-like phenotypes, have replaced established glioblastoma cell lines. However, due to heterogenous tumour growth inherent in glioblastoma, the use of primary cells for orthotopic in vivo studies often requires large experimental group sizes. Therefore, when using intracranial patient-derived xenograft (PDX) approaches, it is advantageous to deploy imaging techniques to monitor tumour growth and allow stratification of mice. Here we show that stable expression of near-infrared fluorescent protein (iRFP) in patient-derived glioblastoma cells enables rapid, direct non-invasive monitoring of tumour development without compromising tumour stemness or tumorigenicity. Moreover, as this approach does not depend on the use of agents like luciferin, which can cause variability due to changing bioavailability, it can be used for quantitative longitudinal monitoring of tumour growth. Notably, we show that this technique also allows quantitative assessment of tumour burden in highly invasive models spreading throughout the brain. Thus, iRFP transduction of primary patient-derived glioblastoma cells is a reliable, cost- and time-effective way to monitor heterogenous orthotopic PDX growth.
Highlights
Despite extensive research, little progress has been made in glioblastoma therapy, owing in part to a lack of adequate preclinical in vivo models to study this disease
We found that infrared fluorescent protein (iRFP)-labelled primary glioblastoma stem-like cells (GSC) retain a comparable level of resistance to established chemotherapeutics
In this paper we show that the use of cell lines stably expressing iRFP overcomes many of these limitations
Summary
Little progress has been made in glioblastoma therapy, owing in part to a lack of adequate preclinical in vivo models to study this disease. We show that stable expression of near-infrared fluorescent protein (iRFP) in patient-derived glioblastoma cells enables rapid, direct non-invasive monitoring of tumour development without compromising tumour stemness or tumorigenicity As this approach does not depend on the use of agents like luciferin, which can cause variability due to changing bioavailability, it can be used for quantitative longitudinal monitoring of tumour growth. It is advantageous to study glioblastoma behaviour in vivo, as brain tissue provides the most appropriate microenvironment for maintenance of the stem signature of patient-derived xenografts (PDX)[7] Due to their intrinsic heterogeneity, orthotopic PDXs display a much greater range of growth rates between animal subjects compared to established glioblastoma cell lines[4,10]. MRIs often only measure indirect indicators of tumour pathology, such as focal brain oedema, which do not necessarily reflect the size and margins of the tumour itself
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