» Articles » PMID: 37147485

Technical Report: Surgical Preparation of Human Brain Tissue for Clinical and Basic Research

Overview
Specialty Neurosurgery
Date 2023 May 5
PMID 37147485
Authors
Affiliations
Soon will be listed here.
Abstract

Background: The study of the distinct structure and function of the human central nervous system, both in healthy and diseased states, is becoming increasingly significant in the field of neuroscience. Typically, cortical and subcortical tissue is discarded during surgeries for tumors and epilepsy. Yet, there is a strong encouragement to utilize this tissue for clinical and basic research in humans. Here, we describe the technical aspects of the microdissection and immediate handling of viable human cortical access tissue for basic and clinical research, highlighting the measures needed to be taken in the operating room to ensure standardized procedures and optimal experimental results.

Methods: In multiple rounds of experiments (n = 36), we developed and refined surgical principles for the removal of cortical access tissue. The specimens were immediately immersed in cold carbogenated N-methyl-D-glucamine-based artificial cerebrospinal fluid for electrophysiology and electron microscopy experiments or specialized hibernation medium for organotypic slice cultures.

Results: The surgical principles of brain tissue microdissection were (1) rapid preparation (<1 min), (2) maintenance of the cortical axis, (3) minimization of mechanical trauma to sample, (4) use of pointed scalpel blade, (5) avoidance of cauterization and blunt preparation, (6) constant irrigation, and (7) retrieval of the sample without the use of forceps or suction. After a single round of introduction to these principles, multiple surgeons adopted the technique for samples with a minimal dimension of 5 mm spanning all cortical layers and subcortical white matter. Small samples (5-7 mm) were ideal for acute slice preparation and electrophysiology. No adverse events from sample resection were observed.

Conclusion: The microdissection technique of human cortical access tissue is safe and easily adoptable into the routine of neurosurgical procedures. The standardized and reliable surgical extraction of human brain tissue lays the foundation for human-to-human translational research on human brain tissue.

Citing Articles

Clinical parameters affect the structure and function of superficial pyramidal neurons in the adult human neocortex.

Lenz M, Kruse P, Eichler A, Straehle J, Hemeling H, Stohr P Brain Commun. 2024; 6(5):fcae351.

PMID: 39474044 PMC: 11518857. DOI: 10.1093/braincomms/fcae351.


A framework for standardised tissue sampling and processing during resection of diffuse intracranial glioma: joint recommendations from four RANO groups.

Karschnia P, Smits M, Reifenberger G, Le Rhun E, Ellingson B, Galldiks N Lancet Oncol. 2023; 24(11):e438-e450.

PMID: 37922934 PMC: 10849105. DOI: 10.1016/S1470-2045(23)00453-9.

References
1.
Andersson M, Avaliani N, Svensson A, Wickham J, Pinborg L, Jespersen B . Optogenetic control of human neurons in organotypic brain cultures. Sci Rep. 2016; 6:24818. PMC: 4838935. DOI: 10.1038/srep24818. View

2.
Jones R, Brito da Silva A, Whittaker R, Woodhall G, Cunningham M . Human brain slices for epilepsy research: Pitfalls, solutions and future challenges. J Neurosci Methods. 2015; 260:221-32. DOI: 10.1016/j.jneumeth.2015.09.021. View

3.
Rojas-Piloni G, Guest J, Egger R, Johnson A, Sakmann B, Oberlaender M . Relationships between structure, in vivo function and long-range axonal target of cortical pyramidal tract neurons. Nat Commun. 2017; 8(1):870. PMC: 5636900. DOI: 10.1038/s41467-017-00971-0. View

4.
Kovacs F, Knop T, Urbanski M, Freiman I, Freiman T, Feuerstein T . Exogenous and endogenous cannabinoids suppress inhibitory neurotransmission in the human neocortex. Neuropsychopharmacology. 2011; 37(5):1104-14. PMC: 3306870. DOI: 10.1038/npp.2011.262. View

5.
Kirov S, Petrak L, Fiala J, Harris K . Dendritic spines disappear with chilling but proliferate excessively upon rewarming of mature hippocampus. Neuroscience. 2004; 127(1):69-80. DOI: 10.1016/j.neuroscience.2004.04.053. View