黑色素细胞与骨髓间充质干细胞复合构建组织工程化皮肤  

Application of Melanocytes and Bone Marrow Mesenchymal Stem Cells in Tissue-Engineered Skin Construction

作　　者：宋可新[1] 乔群[1] 刘大庆[2] 王晓军[1] 赵茹[1] 刘志飞[1] 裴雪涛[2] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院整形外科,北京100032 [2]军事医学科学院野战输血研究所干细胞与再生医学研究室,北京100850

出　　处：《中国医学科学院学报》2011年第4期402-407,483-486,共10页Acta Academiae Medicinae Sinicae

摘　　要：目的探讨构建含有黑色素细胞组织工程化皮肤的方法。方法从人包皮组织中获取黑色素细胞,从人骨髓中获取骨髓间充质干细胞(BMSCs),以二者为种子细胞,按照1∶10的比例混和培养,并与I型胶原膜复合体外构建组织工程化皮肤,对裸鼠创面进行修复。通过大体标本观察、4,6-联脒-2-苯基吲哚(DAPI)体内示踪标记、HE染色、免疫组织化学检测及透射电镜观察等方法,观察组织工程化皮肤修复裸鼠皮肤创面缺损及黑色素细胞的分布情况。结果裸鼠创面皮肤生长良好,DAPI体内示踪标记、免疫组织化学检测S-100蛋白及透射电镜观察可以发现黑色素细胞以正常的组织结构形式分布于创面皮肤。结论黑色素细胞与BMSCs通过适当的比例及体外条件培养,与I型胶原膜复合可以在体外构建出具有黑色素细胞的组织工程化皮肤。Objective To explore the method of constructing tissue-engineered skin using melanocytes and bone marrow mesenchymal stem cells（BMSCs） in vivo.Methods Melanocytes were isolated from human foreskin.BMSCs were isolated from human bone marrow.Both of them were co-cultured at a ratio of 1∶10,and then were implanted into the collagen membrane to construct the tissue-engineered skin,which was applied for wound repair in nude mice.The effectiveness of wound repair and the distribution of melenocytes were evaluated by morphological observation,in vivo 4,6-diamidino-2-phenylindole,dihydrochloride（DAPI） fluorescent staining tracing,HE staining,S-100 immunohistochemistry,and transmission electron microscopy.Results The wounds were satisfactorily repaired among the nude mice.The melenocytes were distributed in the skin with normal structure,as confirmed by DAPI fluorescent staining tracing,HE staining,S-100 immunohistochemistry,and transmission electron microscopy.Conclusion Melanocytes and BMSCs,after proper in vitro culture at an appropriate ratio,can construct the tissue-engineered skin with I type collagen membrane.

关 键 词：黑色素细胞 骨髓间充质干细胞 胶原膜 组织工程化皮肤 

分 类 号：R318.19[医药卫生—生物医学工程]