Title: Single cell multi-omics characterise discrete human tendon cells populations that persist in vitro and on fibrous scaffolds |
Authors: A Gomez-Collignon, R Brown, A Carr, S Dakin, A Lach, C Loizou, M Rogers, R Sharp, A Kendal |
Address: Botnar Research Centre, The Nuffield Department of
Orthopaedics, Rheumatology and Musculoskeletal Sciences, Windmill Road, Oxford, OX3 7LD, UK
|
E-mail: adriankendal at hotmail.com
|
Abstract: Chronic tendinopathy represents a growing healthcare burden in the ageing global population. Curative
therapies remain elusive as the mechanisms that underlie chronic inflammation in tendon disease remain
unclear. Identifying and isolating key pathogenic and reparative cells is essential in developing precision
therapies and implantable materials for improved tendon healing.
Multiple discrete human tendon cell populations have been previously described ex vivo. To determine
if these populations persist in vitro, healthy human hamstring tenocytes were cultured for 8 d on either
tissue culture plastic or aligned electrospun fibres of absorbable polydioxanone. Novel single-cell surface
proteomics combined with unbiased single-cell transcriptomics (CITE-Seq) was used to identify discrete
tenocyte populations.
6 cell populations were found, 4 of which shared key gene expression determinants with ex vivo human
cell clusters: PTX3_PAPPA, POSTN_SCX, DCN_LUM and ITGA7_NES. Surface proteomics found that
PTX3_PAPPA cells were CD10+CD26+CD54+. ITGA7_NES cells were CD146+ and POSTN_SCX cells were
CD90+CD95+CD10+.
Culture on the aligned electrospun fibres favoured 3 cell subtypes (DCN_LUM, POSTN_SCX and PTX3_
PAPPA), promoting high expression of tendon-matrix-associated genes and upregulating gene sets enriched
for TNF-a and IL-6/STAT3 signalling.
Discrete human tendon cell subpopulations persisted in in vitro culture and could be recognised by
specific gene and surface-protein signatures. Aligned polydioxanone fibres promoted the survival of 3
clusters, including pro-inflammatory PTX3-expressing CD10+CD26+CD54+ cells found in chronic tendon
disease. These results improved the understanding of preferred culture conditions for different tenocyte
subpopulations and informed the development of in vitro models of tendon disease.
|
Keywords: Tenocyte, tendon fibroblast, human tendon, single-cell RNA sequencing, CITE-Seq, transcriptomic,
tendinopathy, in vitro, culture, polydioxanone, scaffolds, fibres.
|
Publication date: August 2nd 2022 |
Article download: Pages
1-20 (PDF file)
DOI: 10.22203/eCM.v044a01 |