Department of Otorhinolaryngology, Gangnam Severance Hospital, Yonsei University College of Medicine, 211 Eonju-ro, Gangnam-gu, Seoul, 06273, Republic of Korea.
Stem Cell Res Ther. 2017 Sep 29;8(1):200. doi: 10.1186/s13287-017-0650-y.
Although 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.
Single 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.
Self-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.
These 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.
虽然喉组织固有间充质基质细胞（MSCs）已经被描述过，但它们的独特特征和功能尚未被充分探索。因此，我们研究了从大鼠会厌黏膜（EM）、固有层（LP）和黄斑（MF）中分离的单个克隆群体的干细胞特征和再生潜力，以确定它们是否包含喉组织固有干细胞。
通过对大鼠喉 EM、LP 和 MF 的显微解剖，分离出单个克隆的喉细胞。从三个喉亚区选择并体外扩增多个克隆群体。我们比较了喉 MSC 样细胞的自我更新和分化潜能、细胞表面表型和基因表达谱，以及骨髓间充质干细胞（BM-MSCs）的特征。我们还研究了喉细胞在放射性喉损伤动物模型中的再生潜力。
从大鼠 EM、LP 和 MF 中获得了具有自我更新能力的克隆细胞群体。EM 衍生和 LP 衍生的克隆细胞具有成纤维细胞样特征，而 MF 驻留的克隆细胞具有星状细胞形态和含有维生素 A 的脂滴。所有喉克隆细胞群体均具有 MSC 样细胞表面标志物表达（CD29、CD44、CD73 和 CD90），并具有分化为骨和软骨细胞谱系的潜能；EM 衍生和 MF 衍生的细胞，但不是 LP 衍生的细胞，也能够分化为脂肪细胞。从喉亚区分离的克隆细胞表现出不同的细胞外基质相关基因表达。我们发现，与 BM-MSCs 相比，间充质和星状细胞相关基因结蛋白和巢蛋白在喉 MSC 样细胞中丰富（P < 0.001）。生长分化因子 3（GDF3）和胶质纤维酸性蛋白（GFAP）的转录和蛋白水平在 MF 衍生的细胞中高于其他喉群体（P < 0.001）。移植后 4 周，喉 MF 衍生和 EM 衍生的细胞在辐射损伤后有助于喉上皮和/或腺的再生。
这些结果表明，具有 MSC 特征的细胞群体存在于喉的 EM、LP 和 MF 中。喉 MSC 样细胞有助于损伤后的喉再生；需要进一步研究以阐明不同群体在喉组织再生中的作用，以及它们在治疗喉疾病方面的临床意义。
