Fredriksson Camilla, Kratz Gunnar, Huss Fredrik
Institution of Biomedicine and Surgery, Department of Experimental Plastic Surgery, Faculty of Health Science, Linköpings Universitet, S-581 82 Linköping, Sweden.
Burns. 2008 Mar;34(2):212-9. doi: 10.1016/j.burns.2007.03.008. Epub 2007 Aug 3.
Transplantation of autologous cultured keratinocytes in single cell suspension is useful in the treatment of burns. The reduced time needed for culture, and the fact that keratinocytes in suspension can be transported from the laboratory to the patient in small vials, thus reducing the costs involved and be stored (frozen) in the clinic for transplantation when the wound surfaces are ready, makes it appealing. We found few published data in the literature about actual cell survival after transplantation of keratinocytes in single cell suspension and so did a comparative in vitro study, considering commonly used application techniques. Human primary keratinocytes were transplanted in vitro in a standard manner using different techniques. Keratinocytes were counted before and after transplantation, were subsequently allowed to proliferate, and counted again on days 4, 8, and 14 by vital staining. Cell survival varied, ranging from 47 to >90%, depending on the technique. However, the proliferation assays showed that the differences in numbers diminished after 8 days of culture. Our findings indicate that a great number of cells die during transplantation but that this effect is diminished if cells are allowed to proliferate in an optimal milieu. A burned patient's wounds cannot be regarded as the optimal milieu, and using less harsh methods of transplantation may increase the take rate and wound closing properties of autologous keratinocytes transplanted in a single cell suspension.
将自体培养的角质形成细胞以单细胞悬液形式进行移植，在烧伤治疗中很有用。培养所需时间缩短，且悬液中的角质形成细胞可装在小瓶中从实验室运至患者处，从而降低相关成本，并可在临床储存（冷冻），待创面准备好时进行移植，这些都使其具有吸引力。我们在文献中很少找到关于单细胞悬液形式的角质形成细胞移植后实际细胞存活率的已发表数据，因此进行了一项比较体外研究，考虑了常用的应用技术。人原代角质形成细胞采用不同技术以标准方式进行体外移植。在移植前后对角质形成细胞进行计数，随后使其增殖，并在第4、8和14天通过活细胞染色再次计数。细胞存活率各不相同，根据技术不同，范围在47%至>90%之间。然而，增殖试验表明，培养8天后数量差异减小。我们的研究结果表明，大量细胞在移植过程中死亡，但如果细胞在最佳环境中增殖，这种影响会减小。烧伤患者的创面不能被视为最佳环境，采用不那么苛刻的移植方法可能会提高单细胞悬液形式移植的自体角质形成细胞的成活率和创面闭合性能。
