组织工程皮肤种子细胞生物学特性的研究

目的建立一种分离培齐组织工程皮肤种子细胞的方法，并对细胞的生物学特性进行研究。方法分别通过Dispase-胰蛋白酶消化法和组织块法对表皮干细胞和成纤维细胞进行分离培养，利用倒置相差显微镜和扫描电镜等对其进行形态学观察，MTT比色法对细胞的增殖动态进行测定，免疫组化法对两种细胞进行鉴定。结果分离的表皮细胞和成纤维细胞可在体外快速贴壁生长、增殖，活力较好，免疫组织化学染色为阳性。结论通过Dispase-胰蛋白酶消化法和组织块法分别获得的表皮干细胞和成纤维细胞在体外能够快速扩增、培养，可为下一步组织工程皮肤的构建提供充足、可靠的细胞源。

Biological characteristics of seed cell of tissue engineered skins

Objective To establish the isolation, cultivation technique of human skin seed cells for tissue engineering and study the biological characteristics. Methods The epidermal stem cells and fibroblast were isolated from the human skin by two-step digesting of dispase and trypsin and explant culture method. Cellular morphologies were observed with inverted phase contrast microscopy and scanning electron microscopy. Cell viability was detected with MTT. Cell type was identified with immunohistochemistry. Results The isolated epidermal cells and fibroblasts could rapidly grow and proliferate, which were positive in immunostaining. Conclusions The human skin ceils, including epidermal cells and fibroblasts, acquired with the methods mentioned above could be cultured and proliferated rapidly in vitro, which would provide sufficient and reliable cells resources for tissue engineering skin.