Boranić M, Jakić-Razumović J, Stanović S, Kljenak A, Fattorini I
Institut Ruder Bosković, Zavod za molekularnu medicinu, Laboratorij za eksperimentalnu hematologiju, imunologiju i onkologiju, Zagreb.
Lijec Vjesn. 1999 Apr-May;121(4-5):137-43.
Skin is the largest organ of the human body (8-10 kg, 1.5-2.0 m2, 10(11) cells of epidermal, mesenchymal and neural origin). Although endowed with remarkable regeneration ability, the recovery after major injuries viz. burns requires appropriate surgical treatment, temporary coverage of defects and supportive measures. Large defects are covered with viable transplants of autologous or allogeneic skin, frozen or lyophilized human and animal skin, bioartificial tissues made of synthetic or biodegradable materials, sheets of keratinocytes cultured in vitro. The use of autotransplants is limited by the size of preserved skin areas as well as by general condition of the patient. Allotransplants collected from cadavers or volunteers are rejected after 1 or 2 weeks and thus afford only temporary coverage. Grafts of human or animal skin devitalized by lyophilization or freezing in glycerol accommodate connective tissue and blood vessels ingrowing from the graft bed but eventually dissolve. Artificial skin consists of collagen, chondroitin or similar fiber network (substituting the dermis) covered by semipermeable silicon foil (substituting the epidermis). After healing in, the silicon foil is peeled off and replaced by skin autotransplants or autologous keratinocytes grown and expanded in vitro. The technique for massive production of human keratinocytes, invented some twenty years ago, has been applied for clinical purposes by several specialized centers. During the culture period of approximately three weeks the keratinocyte population may enlarge five to ten thousandfold. Keratinocytes obtained from a 1.5 cm2 piece of skin (half of a postal stamp) may thus yield progeny sufficient for the coverage of 1.5 m2, which is almost the total body surface. The period required for culturing autologous keratinocytes is bridged by temporary transplants and vigorous supportive treatment of the patient. Cultured keratinocytes display all essential features of keratinocytes in situ. They divide and differentiate, express membrane structures required for intercellular communication and reception of signals regulating cell division and differentiation, secrete cytokines. In addition to clinical application, the culture of human keratinocytes is a convenient and useful model for studies of cellular biology. This review is illustrated by first examples of keratinocyte cultures grown in our laboratory.
皮肤是人体最大的器官（8 - 10千克，1.5 - 2.0平方米，由表皮、间充质和神经来源的10¹¹个细胞组成）。尽管皮肤具有显著的再生能力，但遭受重大损伤如烧伤后，其恢复仍需要适当的手术治疗、缺损的临时覆盖以及支持措施。大面积缺损可用自体或异体皮肤的活移植、冷冻或冻干的人及动物皮肤、由合成或可生物降解材料制成的生物人工组织、体外培养的角质形成细胞片覆盖。自体移植的使用受到剩余皮肤面积大小以及患者一般状况的限制。从尸体或志愿者身上获取的异体移植在1至2周后会被排斥，因此仅能提供临时覆盖。经冻干或在甘油中冷冻处理而失去活力的人或动物皮肤移植物，能容纳从移植床向内生长的结缔组织和血管，但最终会溶解。人工皮肤由胶原蛋白、软骨素或类似的纤维网络（替代真皮）组成，上面覆盖着半透性硅箔（替代表皮）。植入后愈合时，硅箔被剥离，代之以自体皮肤移植或体外培养并扩增的自体角质形成细胞。大约二十年前发明的大规模生产人角质形成细胞的技术，已被几个专业中心应用于临床。在大约三周的培养期内，角质形成细胞群体可能扩大5000至10000倍。因此，从一块1.5平方厘米的皮肤（半张邮票大小）获取的角质形成细胞可产生足够覆盖1.5平方米的后代，这几乎是整个身体表面。自体角质形成细胞培养所需的时间通过临时移植和对患者的积极支持治疗来弥补。培养的角质形成细胞展现出原位角质形成细胞的所有基本特征。它们进行分裂和分化，表达细胞间通讯以及接收调节细胞分裂和分化信号所需的膜结构，分泌细胞因子。除了临床应用外，人角质形成细胞培养对于细胞生物学研究也是一个方便且有用的模型。本综述通过我们实验室培养的角质形成细胞的首批实例加以说明。
