@article{b9fe875158014861a2a68e38fa816cf4,
title = "Generation of 3D ex vivo mouse- and patient-derived glioma explant slice model for integration of confocal time-lapse imaging and spatial analysis",
abstract = "Development of spatial-integrative pre-clinical models is needed for glioblastoma, which are heterogenous tumors with poor prognosis. Here, we present an optimized protocol to generate three-dimensional ex vivo explant slice glioma model from orthotopic tumors, genetically engineered mouse models, and fresh patient-derived specimens. We describe a step-by-step workflow for tissue acquisition, dissection, and sectioning of 300-μm tumor slices maintaining cell viability. The explant slice model allows the integration of confocal time-lapse imaging with spatial analysis for studying migration, invasion, and tumor microenvironment, making it a valuable platform for testing effective treatment modalities. For complete details on the use and execution of this protocol, please refer to Comba et al. (2022).1",
keywords = "Biotechnology and Bioengineering, Cancer, Cell Biology, Cell-based Assays, Health Sciences, Microscopy, Model Organisms",
author = "Andrea Comba and Varela, {Maria Luisa} and Faisal, {Syed M.} and Abel, {Clifford C.} and Argento, {Anna E.} and Al-Holou, {Wajd N.} and Hollon, {Todd C.} and Perelman, {Jacqueline D.} and Dunn, {Patrick J.} and Sebastien Motsch and Castro, {Maria G.} and Lowenstein, {Pedro R.}",
note = "Funding Information: We thank all members of our laboratory for advice and comments on this work. This work was supported by National Institutes of Health, National Institute of Neurological Disorders and Stroke (NIH/NINDS) grants R37-NS094804, R01-NS105556, R21-NS107894, R21-NS091555, and R01-NS074387 to M.G.C.; National Institute of Neurological Disorders and Stroke (NIH/NINDS) grants R01-NS076991, R01-NS096756, R01-NS082311, R01-NS122234, and R01-NS127378 to P.R.L.; National Institute of Biomedical Imaging and Bioengineering (NIH/NIBI) R01-EB022563; National Cancer Institute (NIH/NCI) U01CA224160; Rogel Cancer Center at The University of Michigan G023089 to M.G.C.; Ian's Friends Foundation grant G024230; Leah's Happy Hearts Foundation grant G013908; Pediatric Brain Tumor Foundation grant G023387; and ChadTough Foundation grant G023419 to P.R.L. RNA Biomedicine grant F046166 to M.G.C.; and Health and Human Services, National Institutes of Health, UL1 TR002240 to Michigan Institute for Clinical and Health Research (MICHR), Postdoctoral Translational Scholars Program (PTSP), Project F049768 to A.C. Conception of the initial project, A.C. and P.R.L.; design and optimization of the model, A.C.; execution of experiments, A.C. M.L.V. S.M.F. C.C.A. P.J.D. and A.E.A.; optimization and acquisition of time-lapse confocal videos, A.C.; post-imaging statistical and mathematical analysis, S.M.F.; acquisition of data, analysis, and interpretation, A.C. S.M.F. M.L.V. C.C.A. J.D.P. S.M. M.G.C. and P.R.L.; provision of patient-derived glioma specimens, W.N.A.-.H. and T.C.H.; manuscript writing, A.C. M.L.V. M.S.F. C.C.A. A.E.A. and P.R.L.; study supervision, M.G.C. and P.R.L.; all authors reviewed the final version of the manuscript. The authors declare no competing interests. Funding Information: We thank all members of our laboratory for advice and comments on this work. This work was supported by National Institutes of Health , National Institute of Neurological Disorders and Stroke (NIH/NINDS) grants R37-NS094804 , R01-NS105556 , R21-NS107894 , R21-NS091555, and R01-NS074387 to M.G.C.; National Institute of Neurological Disorders and Stroke (NIH/NINDS) grants R01-NS076991 , R01-NS096756 , R01-NS082311 , R01-NS122234 , and R01-NS127378 to P.R.L.; National Institute of Biomedical Imaging and Bioengineering (NIH/NIBI) R01-EB022563 ; National Cancer Institute (NIH/NCI) U01CA224160 ; Rogel Cancer Center at The University of Michigan G023089 to M.G.C.; Ian{\textquoteright}s Friends Foundation grant G024230; Leah{\textquoteright}s Happy Hearts Foundation grant G013908; Pediatric Brain Tumor Foundation grant G023387; and ChadTough Foundation grant G023419 to P.R.L. RNA Biomedicine grant F046166 to M.G.C.; and Health and Human Services , National Institutes of Health, UL1 TR002240 to Michigan Institute for Clinical and Health Research (MICHR), Postdoctoral Translational Scholars Program (PTSP), Project F049768 to A.C. Publisher Copyright: {\textcopyright} 2023 The Author(s)",
year = "2023",
month = jun,
day = "16",
doi = "10.1016/j.xpro.2023.102174",
language = "English (US)",
volume = "4",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "2",
}