Dissociated neurons and glial cells derived from rat inferior colliculi after digestion with papain

Odett Kaiser, Pooyan Aliuos, Kirsten Wissel, Thomas Lenarz, Darja Werner, Günter Reuter, Andrej Kral, Athanasia Warnecke

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The formation of gliosis around implant electrodes for deep brain stimulation impairs electrode-tissue interaction. Unspecific growth of glial tissue around the electrodes can be hindered by altering physicochemical material properties. However, in vitro screening of neural tissue-material interaction requires an adequate cell culture system. No adequate model for cells dissociated from the inferior colliculus (IC) has been described and was thus the aim of this study. Therefore, IC were isolated from neonatal rats (P3-5) and a dissociated cell culture was established. In screening experiments using four dissociation methods (Neural Tissue Dissociation Kit [NTDK] T, NTDK P; NTDK PN, and a validated protocol for the dissociation of spiral ganglion neurons [SGN]), the optimal media, and seeding densities were identified. Thereafter, a dissociation protocol containing only the proteolytic enzymes of interest (trypsin or papain) was tested. For analysis, cells were fixed and immunolabeled using glial- and neuron-specific antibodies. Adhesion and survival of dissociated neurons and glial cells isolated from the IC were demonstrated in all experimental settings. Hence, preservation of type-specific cytoarchitecture with sufficient neuronal networks only occurred in cultures dissociated with NTDK P, NTDK PN, and fresh prepared papain solution. However, cultures obtained after dissociation with papain, seeded at a density of 2×104 cells/well and cultivated with Neuro Medium for 6 days reliably revealed the highest neuronal yield with excellent cytoarchitecture of neurons and glial cells. The herein described dissociated culture can be utilized as in vitro model to screen interactions between cells of the IC and surface modifications of the electrode.

Original languageEnglish
Article numbere80490
Pages (from-to)1-14
Number of pages14
JournalPLoS ONE
Volume8
Issue number12
DOIs
Publication statusPublished - 12 Dec 2013
Externally publishedYes

Fingerprint Dive into the research topics of 'Dissociated neurons and glial cells derived from rat inferior colliculi after digestion with papain'. Together they form a unique fingerprint.

Cite this