大鼠颌突外胚间充质细胞向骨骼肌细胞诱导分化和三维培养观察  

Differentiation of ectomesenchymal cells of fetal rat mandibular process to skeleton muscle cells by DMSO and observed in three-dimension condition

作　　者：姚远 聂鑫[2] 金岩[2] 

机构地区：[1]珠海市口腔医院,广东珠海519000 [2]第四军医大学口腔医学院组织病理教研室

出　　处：《牙体牙髓牙周病学杂志》2006年第6期307-310,共4页Chinese Journal of Conservative Dentistry

基　　金：珠海市科技计划项目(珠科2002-37号)

摘　　要：目的:探讨体外诱导外胚间充质细胞向骨骼肌细胞分化的潜能,利用分化细胞构建组织工程骨骼肌模型。方法:取妊娠10.5 d胎鼠颌突外胚间充质细胞,纯化后分别在含1、5、10 mL/L体积浓度二甲基亚砜的DMEM/F12培养基中培养。相差显微镜下观察细胞的形态变化,成肌相关因子免疫组化鉴定细胞性质,将诱导后的细胞种植于胶原膜,并于培养3、6 d后用光镜、扫描电镜观察细胞的生长情况。结果:培养3 d后,外胚间充质细胞形态出现成肌样细胞变化,细胞变得大而扁平,培养5 d后细胞出现管样结构。免疫组化鉴定显示:添加二甲基亚砜后,大部分细胞出现成肌相关因子的表达,其中5 mL/L实验组效果最好,约75%的细胞转化为骨骼肌细胞。对照细胞形态未见明显改变,免疫组化鉴定未能诱导分化出骨骼肌细胞。种植于BAM膜后,细胞生长良好,并可形成复层结构。结论:二甲基亚砜可诱导外胚间充质细胞向骨骼肌细胞分化,最佳浓度为5 mL/L;在BAM膜上分化细胞生长状态良好,说明该生物可降解膜是构建组织工程骨骼肌的适合材料。AIM：To investigate the ability of dimethyl sulphoxide（DMSO） on inducing the ectomesenchymal cells to skeleton muscle cells and using these cell to construct the three - dimension model. METHODS： Different concentrations of DMSO were added to the third passages of ectomesenchymal cells. The morphology was observed by phase - contrast microscopy. Immunohistochemistry was used to identify the characteristics of cells. The three - dimension model was constructed with collagen. RESULTS ： There were great changes in the morphology of the cells treated by DMSO. The ceils changed from fibroblast - like to big and flat. After culture for 5 days, appeared tube structure. After treated with DMSO, the samples were studied by immunohistochemistry. About 75% cells in the 5mL/L experimental group began expression muscle relative factors which means these cell transform into skeleton type. Differentiated cells implanted into BAM membrane could form muhilayer structure. CONCLUSION： The ectomesenchymal cells could be differentiated to skeleton muscle ceils by DMSO, and the collagen membrane were the ideal materials in construction of tissue - engineering skeleton muscle.

关 键 词：外胚间充质细胞 骨骼肌细胞 组织工程 二甲基亚砜 

分 类 号：R780.2[医药卫生—口腔医学]