Kumai Yoshihiko, Kobler James B, Park Hyoungshin, Lopez-Guerra Gerardo, Karajanagi Sandeep, Herrera Victoria L M, Zeitels Steven M
Department of Surgery, Harvard Medical School, Center for Laryngeal Surgery and Voice Rehabilitation, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Laryngoscope. 2009 Apr;119(4):799-805. doi: 10.1002/lary.20149.
OBJECTIVES/HYPOTHESIS: To explore adipose-derived stem cell/fibroblast interactions as a potential remodeling pathway for vocal fold scar. STUDY DESIGN: Fibroblasts and adipose-derived stem/stromal cells (ASCs) were cultured alone and in combination in a cell-contact-independent paracrine system. Analyses of cell proliferation, and the production of hyaluronic acid (HA) and collagen were performed on samples collected on days 1, 3, and 7. METHODS: Normal fibroblasts (NFs) were isolated bilaterally from the subepithelial lamina propria of two normal ferret vocal folds. Scar fibroblasts (SFs) were isolated from vocal folds that were electrocauterized 2 weeks before harvest. ASCs were isolated from lipoaspirated subcutaneous abdominal fat of two ferrets. A transwell cell-contact-independent cell communication culturing system was used for coculture experiments. Cells were seeded at 50,000/well in both monoculture and coculture experiments. RESULTS: In monoculture, SFs proliferated faster and produced less HA and more collagen than NFs at day 7 (P < .05). In SF/ASC coculture, SF proliferation was diminished and collagen production at day 7 decreased (P < .05). HA production did not differ between monoculture and coculture conditions. CONCLUSIONS: Normal and scar-tissue-derived vocal fold fibroblasts maintain phenotypic differences in culture, thus validating this in vitro scar model. In co-culture, contact-independent crosstalk occurs between SFs and ASCa, leading to less collagen secretion. The data support the hypothesis that ASCs can induce favorable remodeling of scarred vocal folds in vivo by their interactions with endogenous fibroblasts.
目的/假设:探讨脂肪来源干细胞与成纤维细胞的相互作用,作为声带瘢痕潜在的重塑途径。 研究设计:在细胞非接触旁分泌系统中单独培养及联合培养成纤维细胞和脂肪来源的干/基质细胞(ASC)。在第1、3和7天收集的样本上进行细胞增殖分析以及透明质酸(HA)和胶原蛋白生成分析。 方法:从两只正常雪貂声带的上皮下固有层双侧分离出正常成纤维细胞(NF)。瘢痕成纤维细胞(SF)从收获前2周进行电灼的声带中分离。从两只雪貂的腹部抽脂皮下脂肪中分离出ASC。采用Transwell细胞非接触细胞通讯培养系统进行共培养实验。在单培养和共培养实验中,细胞均以50,000个/孔接种。 结果:在单培养中,第7天时SF的增殖速度比NF更快,且HA生成更少,胶原蛋白生成更多(P < 0.05)。在SF/ASC共培养中,SF的增殖减少,第7天时胶原蛋白生成下降(P < 0.05)。单培养和共培养条件下HA生成无差异。 结论:正常和瘢痕组织来源的声带成纤维细胞在培养中保持表型差异,从而验证了这个体外瘢痕模型。在共培养中,SF和ASC之间发生非接触性串扰,导致胶原蛋白分泌减少。数据支持以下假设:ASC可通过与内源性成纤维细胞相互作用,在体内诱导瘢痕化声带发生有利的重塑。
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