Human organotypic brain slice culture: a novel framework for environmental research in neuro-oncology

Abstract

When it comes to the human brain, models that closely mimic in vivo conditions are lacking. Living neuronal tissue is the closest representation of the in vivo human brain outside of a living person. Here, we present a method that can be used to maintain therapeutically resected healthy neuronal tissue for prolonged periods without any discernible changes in tissue vitality, evidenced by immunohistochemistry, genetic expression, and electrophysiology. This method was then used to assess glioblastoma (GBM) progression in its natural environment by microinjection of patient-derived tumor cells into cultured sections. The result closely resembles the pattern of de novo tumor growth and invasion, drug therapy response, and cytokine environment. Reactive transformation of astrocytes, as an example of the cellular nonmalignant tumor environment, can be accurately simulated with transcriptional differences similar to those of astrocytes isolated from acute GBM specimens. In a nutshell, we present a simple method to study GBM in its physiological environment, from which valuable insights can be gained. This technique can lead to further advancements in neuroscience, neuro-oncology, and pharmacotherapy.

Spatiotemporal heterogeneity of glioblastoma is dictated by microenvironmental interference

“…Human neocortical slices were prepared as recently described 21,55 . Resected cortical tissue (assessed by EEG and MRI) was immediately brought to the lab in the “preparation medium” (Gibco Hibernate™ media supplemented with 1 mM Gibco GlutaMax™, 13 mM Glucose, 30 mM NMDG and 1% Anti-Anti) saturated with carbogen (95% O2 and 5% CO2).…”

Crosstalk between lymphoid and myeloid cells orchestrates glioblastoma immunity through Interleukin 10 signaling

“…To validate our computational findings, we made use of the recently described human neocortical GBM model, where the cellular architecture of the CNS is well preserved 15,25 . We cultured non-infiltrated neocortical slices (defined in a recent report 15,25 ) coupled with autografted T cells along with myeloid cell depletion, to understand the communication between myeloid cells in the tumor microenvironment along with lymphoid cells. Three days after chemical depletion of the myeloid cells, we injected a primary cell line (BTSC#233, GFP-tagged, previously characterized by RNA-seq profiling as mesenchymal) 26 .…”

Practical Review on Preclinical Human 3D Glioblastoma Models: Advances and Challenges for Clinical Translation

“…To establish this model, healthy brains are needed and mainly mouse and rat brains are used for this purpose ( Figure 3 ). Alternatively, Heiland and collaborators used human healthy brain coming from the periphery of the tumor [ 82 , 83 ]. After extraction of the rodent brain out of the skull, the brain is cut following the coronal axis, into slices of 200 to 400 µm thickness with a vibratome or a tissue chopper.…”

“…It is possible to embed the brain into low melting agarose [ 84 ] or to directly stick it to the platform before cutting [ 85 ]. During slicing, it is recommended to bubble a 95% O 2 /5% CO 2 gas mixture into the vibratome reservoir with cold Phosphate Buffered Saline (PBS), neurobasal medium or hibernate-A medium supplemented with antibiotics/antimycotics [ 82 , 83 , 84 , 86 ]. Slices are then transferred into 6-well plates on a cell culture insert [ 82 , 83 , 84 ].…”