Comparative characteristics of laryngeal-resident mesenchymal stromal cell populations isolated from distinct sites in the rat larynx

Abstract

BackgroundAlthough tissue-resident mesenchymal stromal cells (MSCs) in the larynx have been described, their distinct characteristics and roles have not been thoroughly explored. Therefore, we investigated stem cell characteristics and regenerative potentials of single clonal populations isolated from rat epiglottic mucosa (EM), lamina propria (LP), and macula flava (MF) to determine whether they comprised laryngeal tissue-resident stem cells.MethodsSingle clonal laryngeal cells were isolated following microdissection of the EM, LP, and MF from the rat larynx. Several clonal populations from the three laryngeal subsites were selected and expanded in vitro. We compared the stem cell characteristics of self-renewal and differentiation potential, as well as the cell surface phenotypes and gene expression profiles, of laryngeal MSC-like cells to that of bone marrow MSCs (BM-MSCs). We also investigated the regenerative potential of the laryngeal cells in a radiation-induced laryngeal injury animal model.ResultsSelf-renewing, clonal cell populations were obtained from rat EM, LP, and MF. EM-derived and LP-derived clonal cells had fibroblast-like features, while MF-resident clonal cells had stellate cell morphology and lipid droplets containing vitamin A. All laryngeal clonal cell populations had MSC-like cell surface marker expression (CD29, CD44, CD73, and CD90) and the potential to differentiate into bone and cartilage cell lineages; EM-derived and MF-derived cells, but not LP-derived cells, were also able to differentiate into adipocytes. Clonal cells isolated from the laryngeal subsites exhibited differential extracellular matrix-related gene expression. We found that the mesenchymal and stellate cell-related genes desmin and nestin were enriched in laryngeal MSC-like cells relative to BM-MSCs (P < 0.001). Growth differentiation factor 3 (GDF3) and glial fibrillary acidic protein (GFAP) transcript and protein levels were higher in MF-derived cells than in other laryngeal populations (P < 0.001). At 4 weeks after transplantation, laryngeal MF-derived and EM-derived cells contributed to laryngeal epithelial and/or glandular regeneration in response to radiation injury.ConclusionsThese results suggest that cell populations with MSC characteristics reside in the EM, LP, and MF of the larynx. Laryngeal MSC-like cells contribute to regeneration of the larynx following injury; further investigation is needed to clarify the differential roles of the populations in laryngeal tissue regeneration, as well as the clinical implications for the treatment of laryngeal disease.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0650-y) contains supplementary material, which is available to authorized users.

Characteristics and Responses of Human Vocal Fold Cells in a Vibrational Culture Model

“…In addition, previous studies on stellate cells of the liver and pancreas have reported that tissue‐resident stem cell differentiation contributed to tissue regeneration of injured organs . A recent study has also implied that MFe‐derived stellate cells possesses regenerative potential of vocal fold tissues . Thus, further comparative study on both human VF fibroblasts (hVFFs) and human MF‐SCs (hMF‐SCs) is required to understand their physiological properties and roles in VF tissues.…”

“…9,10 A recent study has also implied that MFe-derived stellate cells possesses regenerative potential of vocal fold tissues. 11 Thus, further comparative study on both human VF fibroblasts (hVFFs) and human MF-SCs (hMF-SCs) is required to understand their physiological properties and roles in VF tissues.…”