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Int J Mol Sci. 2018 Jun 13;19(6). pii: E1761. doi: 10.3390/ijms19061761.

Quality Assessment of Surgical Disc Samples Discriminates Human Annulus Fibrosus and Nucleus Pulposus on Tissue and Molecular Level.

Author information

1
Charité-Universitätsmedizin Berlin, Freie Universität Berlin, Humboldt-Universität zu Berlin and Berlin Institute of Health, Tissue Engineering Laboratory and Berlin-Brandenburg Center for Regenerative Therapies, 13353 Berlin, Germany. ann-kathrin.schubert@charite.de.
2
CO.DON AG, 14513 Teltow, Germany. ann-kathrin.schubert@charite.de.
3
CO.DON AG, 14513 Teltow, Germany. jsmink@gmx.de.
4
Department of Neurosurgery, University Medical Center Mannheim, Heidelberg University, 68167 Mannheim, Germany. Mirko.Arp@umm.de.
5
Charité-Universitätsmedizin Berlin, Freie Universität Berlin, Humboldt-Universität zu Berlin and Berlin Institute of Health, Tissue Engineering Laboratory and Berlin-Brandenburg Center for Regenerative Therapies, 13353 Berlin, Germany. jochen.ringe@charite.de.
6
Department of Neurosurgery, University Medical Center Mannheim, Heidelberg University, 68167 Mannheim, Germany. aldemar.hegewald@medma.uni-heidelberg.de.
7
Department of Neurosurgery and Spine Surgery, Helios Baltic Sea Hospital Damp, 24351 Damp, Germany. aldemar.hegewald@medma.uni-heidelberg.de.
8
Charité-Universitätsmedizin Berlin, Freie Universität Berlin, Humboldt-Universität zu Berlin and Berlin Institute of Health, Tissue Engineering Laboratory and Berlin-Brandenburg Center for Regenerative Therapies, 13353 Berlin, Germany. michael.sittinger@charite.de.

Abstract

A discrimination of the highly specialised annulus fibrosus (AF) and nucleus pulposus (NP) cells in the mature human intervertebral disc (IVD) is thus far still not possible in a reliable way. The aim of this study was to identify molecular markers that distinguish AF and NP cells in human disc tissue using microarray analysis as a screening tool. AF and NP samples were obtained from 28 cervical discs. First, all samples underwent quality sorting using two novel scoring systems for small-sized disc tissue samples including macroscopic, haptic and histological evaluation. Subsequently, samples with clear disc characteristics of either AF or NP that were free from impurities of foreign tissue (IVD score) and with low signs of disc degeneration on cellular level (DD score) were selected for GeneChip analysis (HGU1332P). The 11 AF and 9 NP samples showed distinctly different genome-wide transcriptomes. The majority of differentially expressed genes (DEGs) could be specifically assigned to the AF, whereas no DEG was exclusively expressed in the NP. Nevertheless, we identified 11 novel marker genes that clearly distinguished AF and NP, as confirmed by quantitative gene expression analysis. The novel established scoring systems and molecular markers showed the identity of AF and NP in disc starting material and are thus of great importance in the quality assurance of cell-based therapeutics in regenerative treatment of disc degeneration.

KEYWORDS:

annulus fibrosus; marker; microarray analysis; nucleus pulpous; quality assessment

PMID:
29899321
PMCID:
PMC6032144
DOI:
10.3390/ijms19061761
[Indexed for MEDLINE]
Free PMC Article

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