以胶原海绵为载体培养的人表皮细胞移植

目的：将体外培养的人表皮细胞接种到胶原海绵上，构建一种表皮替代物，移植到鼠创面后研究观察皮肤的再生，方法：将手术切下的包皮经中性蛋白酶分离表皮，真皮，再经胰蛋白酶消化制成表皮细胞悬液，移入培养瓶中培养，将处于对数生长期的人表皮细胞直接接种到培养皿中的胶原海绵 ，再加上表皮细胞液培养3天后，移植到裸鼠全层皮肤缺损的创面上，以单纯无接种细胞的胶原海绵作为对照，并进行组织学，免疫组织化学及电子显微镜观察。结果：这种表皮替代物移植到创面后，表皮细胞继续增殖分化，形成一层新生表皮，与对照组相比，创面闭合早且收缩程度小，表皮成熟早且分层较多，基底膜形成较早，表皮下胶原纤维较少。结论：体外培养的表皮细胞能在胶原海绵上生长，移植到创面上后，该细胞可自动移行到创面并形成多层表皮结构，抑制创面收缩，可用于修复皮肤缺损。

TRANSPLANTATION OF CULTURED HUMAN KERATINOCYTE ON COLLAGEN SPONGE

OBJECTIVE: To investigate the skin regeneration using cultured human keratinocytes with collagen sponge transplanted into thickness wound of nude mice. METHODS: Human foreskin from foreskin ectomy procedures was detached with 0.5% Dispase II. Epidermis sheets were separated from dermis and digested with 0.05% Trypsin into single cell suspension. Keratinocytes were cultured and seeded into collagen sponge during logarithmic growth phase. After 3 days, the keratinocytes-collagen sponge were grafted on full thickness wound of nude mice, compared with simple collagen sponge without keratinocytes. The histological, immunohistochemical examination and electron microscopy were detected. RESULTS: After the epidermal substitute was grafted onto wound, the human keratinocytes were able to further proliferate and differentiate and develop into new epithelia. Compared with the control group, the wound healed earlier and contracted less, epithelia matured earlier, and the collagen fiber was less beneath epithelia. CONCLUSION: Keratinocytes can grow on collagen sponge and migrate onto wound to develop into stratified epithelia and inhibit wound contract. The keratinocyte graft can be used to repair skin defect.