The Department of Biomedical and Clinical Sciences (BKV), Linköping University Hospital, 401A, Building 462, Floor 11, P.O. Box 581 85, Linköping, Sweden.
Department of Hand Surgery and Plastic Surgery and Burns, Linköping University Hospital, Linköping, Sweden.
Sci Rep. 2020 Sep 2;10(1):14486. doi: 10.1038/s41598-020-71553-2.
The rules governing Medicinal Products in the European Union necessitates the production of cell-based therapy in good manufacturing practice facilities. The produced cells may need several hours in transportation to reach the application sites. In this study, we investigated four candidate solutions for transporting human keratinocytes. The solutions are (1) normal saline, (2) saline with 2.5% human serum albumin (Saline + HSA), (3) chemically defined, xeno-free keratinocyte media and (4) keratinocyte media with pituitary bovine extract (PBE-media). One million keratinocytes from three donors were suspended in each solution and kept at 4 °C for up to 24 h. Cells kept in Saline + HSA showed higher viability after 1, 3 and 24 h. Then, equal number of viable cells were seeded on collagenous matrix and cultured for 48 h. The adhesion and colonization were higher in the cells kept in PBE-media, while the keratinocyte surface marker, cytokeratin 14, was present in all studied groups. These results confirmed the suitability of Saline + HSA as a cell transportation solution for clinical use, which will be the choice for the planned clinical trial. Keratinocyte PBE-media can be an alternative for cells transported for research purpose, if the same media type is going to be used in the following experiments.
在欧盟，药品监管法规要求细胞疗法产品在符合良好生产规范的设施中生产。所生产的细胞在运输到应用部位的过程中可能需要数小时。在这项研究中，我们研究了四种用于运输人角质形成细胞的候选解决方案。这些解决方案是：（1）生理盐水，（2）含 2.5%人血清白蛋白的生理盐水（Saline+HSA），（3）化学成分确定、无动物源的角质形成细胞培养基，和（4）含牛垂体提取物的角质形成细胞培养基（PBE 培养基）。将来自三个供体的一百万个人角质形成细胞悬浮在每种溶液中，并在 4°C 下保存长达 24 小时。在 1、3 和 24 小时后，保存在 Saline+HSA 中的细胞显示出更高的活力。然后，将等量的存活细胞接种在胶原蛋白基质上，并培养 48 小时。在 PBE 培养基中保存的细胞的粘附和定植更高，而所有研究组均存在角质形成细胞表面标志物角蛋白 14。这些结果证实了 Saline+HSA 作为临床使用的细胞运输溶液的适用性，这将是计划中的临床试验的选择。如果在后续实验中使用相同的培养基类型，那么 PBE 培养基可以作为用于研究目的运输细胞的替代选择。
