含表皮细胞的无细胞真皮复合皮的构建及生长活性研究

为探讨体外构建复合皮的可能性，制备来源于猪皮的无细胞真皮基质，在其表皮面种植表皮细胞于体外培养，定期消化分离无细菌真皮上的表皮细胞，采用细胞计数法和增殖试验观察表皮细胞增殖活力，并分别于1周和2周后通过组织切片HE染色、扫描电镜观察细胞生长状况。结果显示，表皮细胞随培养时间延长，数量明显增多，且仍具有一定的增殖潜力。培养1周、2周后分别形成单层细胞膜片及含3－6层细胞的复层膜片。提示以异种无细菌真皮为载体，可于体外成功构建含表皮细胞层的活性复合皮。

CONSTRUCTION AND GROWTH ABILITY STUDY OF A COMPOSITE SKIN COMPOSED OF KERATINOCYTES AND ACELLULAR DERMAL MATRIX

To investigate the possibility of constructing composite skin, keratinocytes were cultivated in vitro on the epidermal surface of cell free dermis prepared from pig skin.Keratinocytes grown on the dermal matrix were released at selected time points, followed by determining the proliferative capacity with cell number quantity and cell proliferation test. Cells attaching to the dermal matrix after it were seeded for 1 and 2 weeks were observed with histological section HE staining and electron microscopy scanning. Results showed that the number of keratinocytes was markedly increased with culture time. They maintained their proliferative potential after they were seeded on acellular xeno dermal matrix and reached a confluent monolayer or 3 to 6 layers at the 1st and 2nd week after seeding. The data showed that a living composite skin combined with keratinocytes and acellular dermal matrix could be successfully prepared in vitro.