Xie Ju Lin, Li Tiang Zeng, Qi Shao Hai, Huang Bin, Chen Xi Gu, Chen Jiang Ding
Department of Burns, First Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510080, China.
J Plast Reconstr Aesthet Surg. 2007;60(9):983-90. doi: 10.1016/j.bjps.2005.12.062. Epub 2006 Jul 7.
To study and explore the feasibility of using candidate epidermal stem cells with reconstruct tissue-engineered skin for a skin defect.
After the candidate epidermal stem cells were selected directly by rapid adhesion to type IV collagen within 10min from keratinocytes isolated from foreskin epidermis, the TES was constructed by seeding large-scale cultured candidate epidermal stem cells onto a fibroblast-containing dermal substrate, then grafted onto athymic immunodeficient mice with full-thickness skin defects. All specimens were harvested after 1 week, 2 weeks and 4 weeks of transplantation to evaluate by gross, histological, transmission electron microscopic and immunohistochemical techniques its potential to reconstitute a full-thickness skin defect.
The transplanted skin developed a well-differentiated epidermis composed of stratum basale, prickle cell layer, granular layer and stratum corneum and clearly defined dermis with the morphological features of intact skin. The continuous and integral basement membrane zone (BMZ) was established; hemidesmosomes, basal lamina and anchoring fibrils were detected. In the dermis, the collagen of dermal substitute degraded gradually and fibroblasts were aligned in order; lymphocytes, organelle debris, differentiated microvasculature and hyperactive collagen fibrillogenesis were observed. The immunohistochemistry suggested that the keratinocytes of the TES were originated from the human candidate epidermal stem cells and not from the mice. The constructed TES was similar to the uninjured skin in morphological features, which suggested the constructed TES by combining cultured candidate epidermal stem cells with the dermal substrate could satisfy the need for the restoration of skin defects.
研究和探索利用候选表皮干细胞构建组织工程皮肤修复皮肤缺损的可行性。
从包皮表皮分离的角质形成细胞中，通过在10分钟内快速黏附于IV型胶原直接筛选出候选表皮干细胞，将大规模培养的候选表皮干细胞接种到含成纤维细胞的真皮基质上构建组织工程皮肤，然后移植到有全层皮肤缺损的无胸腺免疫缺陷小鼠身上。移植后1周、2周和4周取所有标本，通过大体观察、组织学、透射电子显微镜和免疫组织化学技术评估其修复全层皮肤缺损的潜力。
移植的皮肤形成了由基底层、棘细胞层、颗粒层和角质层组成的分化良好的表皮，以及形态特征与完整皮肤相似的真皮，其界限清晰。建立了连续完整的基底膜带（BMZ）；检测到半桥粒、基膜和锚定纤维。在真皮中，真皮替代物的胶原逐渐降解，成纤维细胞排列有序；观察到淋巴细胞、细胞器碎片、分化的微血管和活跃的胶原纤维形成。免疫组织化学显示，组织工程皮肤的角质形成细胞来源于人候选表皮干细胞而非小鼠。构建的组织工程皮肤在形态特征上与未损伤皮肤相似，这表明将培养的候选表皮干细胞与真皮基质结合构建的组织工程皮肤能够满足皮肤缺损修复的需求。
