中文摘要：

目的探讨喉部黏膜固有层中是否存在间充质干细胞（mesenchymal stem cells，MSC），并对其进行分离培养及鉴定。方法从喉部手术患者切下的组织中获取正常喉黏膜，消化培养法获得固有层来源的细胞，绘制细胞生长曲线观察其增殖情况，采用平板克隆形成实验获得多个不同形态的细胞克隆并计算其克隆形成能力。利用流式细胞仪对培养的第4代细胞进行表面标志物的分析；采用成脂、成骨、成神经诱导液对其进行多项分化能力的研究。结果经上述方法获得的喉黏膜MSC具有较快的增殖能力和较高的克隆形成能力（克隆形成率为7．1％）；高表达间充质的表面标志物CD29（16．9％）、CD44（97．4％）、CD90（89．5％）、CD105（85．9％）、CD146（2．5％）、stro-1（20．4％），不表达造血系的表面标志物CD34（1．2％）、CD45（0．8％）。在体外分别经成脂、成骨和成神经诱导分化后，油红O染色阳性，茜素红染色阳性，神经细胞特异性抗原S100呈阳性表达，提示喉黏膜MSC可向脂肪、骨骼和神经细胞分化。结论喉黏膜固有层中可分离得到具有快速增殖能力及多向分化潜能的MSC。

英文摘要：

Objective To investigate the presence of mesenchymal stem cells （MSC） in laryngeal mucosa, and to seek for the method to isolate, cultivate and identify these cells. Methods Normal laryngeal mueosa was obtained from patients with laryngeal carcinoma during surgery, and the generating mesenehymal cells was obtained by digestive method. The cell growth curve was evaluated by 3-（4. 5- methyhhiozol-2yl）-2.5-diphenyhetrazolium bromide （MTT） assay. Colony forming cell assay was used to screen different morphologic colonies and evaluate clone formation ability. Flow cytometry was performed for the expression of the cells of the 4th passage surface marker profiles. Multiple differentiation potentials were confirmed by adipogenic, osteogenic and neural lineages induction. Results MTT assay and colony forming cell assay showed that laryngeal mucosa MSC had a relatively rapid proliferation capacity and a relatively high clone formation capacity （clone formation rate 7.1% ）. Flow cytometry analysis revealed that the laryngeal mueosa MSC were positive for CD29 （ 16. 9% ）, CD44 （ 97.4% ）, CD90 （ 89. 5% ）, CD105 （ 85.9% ）, CD146 （2. 5% ） and stro-1 （ 20. 4% ）, but negative for CD34 （ 1.2% ） and CD45 （0. 8% ）. Laryngeal mucosa MSC undergone adipogenic, osteogenic and neural lineages induction were positive for oil red staining, alizarin red staining and SIO0 staining respectively, which suggested that laryngeal mucosa MSC could differentiate into adipogenic, osteogenic and neural lineages. Conclusion This study demonstrated that MSC with rapid proliferative capacity and multiple differentiation potential could be obtained from lamina propria of laryngeal mucosa.