Hecht J, Hoefter E A, Hecht J, Haraida S, Nerlich A, Hartinger A, Mühlbauer W, Dimoudis N
Abteilung für Plastische Chirurgie, Städtisches Krankenhaus Bogenhausen, München.
Handchir Mikrochir Plast Chir. 1997 Mar;29(2):101-6.
Epidermal grafts from confluently cultivated keratinocytes have been used since the early eighties for the treatment of severe burns, where the shortage of donor sites for split-thickness skin grafts did not allow for adequate wound coverage. The difficult handling of these grafts as well as the advanced differentiation of their epithelial cells into a multilayer sheet poses a problem for their clinical application. The aim of the study was to characterize cultivated keratinocytes, as well as to observe their migration and proliferation from the MC onto a surface. Keratinocytes were isolated from human foreskin and cultivated in serum-free and serum-containing medium according to a modified method by Rheinwald and Green. Collagen-coated Dextran beads were used as MC. The MC were colonized with keratinocytes using the Spinner culture technique. After seeding the colonized MC into culture flasks, their migration and proliferation was monitored regularly through immunohistochemical studies and measurement of the metabolic cell activity. Immunohistological staining proved that the cells isolated from human foreskin represent keratinocytes of the basal type. Keratinocytes, cultivated with serum-containing and serum free medium, both adhered to the surface of the MC, then migrated onto the surface of the flasks and proliferated to form a multilayer of epithelial cells. In the long-term, a flexible epithelial graft consisting of poorly differentiated keratinocytes should be available, which is simple to produce and easy to handle. This would be an alternative method for treating wounds, where the conventional multilayer epithelial graft (ET) is insufficient.
自20世纪80年代初以来，源自汇合培养角质形成细胞的表皮移植物就被用于治疗严重烧伤，在这种情况下，供体部位不足，无法进行足够的中厚皮片移植以覆盖伤口。这些移植物处理困难，且其上皮细胞高度分化形成多层片层，这给它们的临床应用带来了问题。本研究的目的是对培养的角质形成细胞进行表征，并观察它们从微载体迁移到表面以及在表面增殖的情况。从人包皮中分离角质形成细胞，并根据Rheinwald和Green改良的方法，在无血清和含血清培养基中培养。胶原包被的葡聚糖珠用作微载体。使用旋转培养技术将角质形成细胞接种到微载体上。将接种了角质形成细胞的微载体接种到培养瓶中后，通过免疫组织化学研究和代谢细胞活性测量定期监测它们的迁移和增殖。免疫组织化学染色证明，从人包皮分离的细胞代表基底型角质形成细胞。在含血清和无血清培养基中培养的角质形成细胞均附着在微载体表面，然后迁移到培养瓶表面并增殖形成多层上皮细胞。从长远来看，应该可以获得一种由低分化角质形成细胞组成的柔性上皮移植物，其制备简单且易于操作。这将是治疗传统多层上皮移植物（ET）不足的伤口的一种替代方法。
