The McComb Foundation, Royal Perth Hospital, Perth, WA, Australia.
Burns. 2012 Feb;38(1):44-51. doi: 10.1016/j.burns.2011.03.001. Epub 2011 Nov 12.
The use of non-cultured autologous cells to promote wound healing and in reconstructive procedures is increasing. One common method for preparing these cells is the use of the ReCell(®) device. However, despite its current clinical use, no characterisation of the cell suspension produced using a ReCell(®) device has been published.
To characterise the ReCell suspension that is applied to wounds for cell type, viability, yield, stability and proliferative potential.
The ReCell(®) device was used to harvest cells from a 2 cm(2) piece of split-thickness skin isolated using a dermatome. The resulting cell suspension was analysed for cell yield, cell type, viability over time, proliferative potential and reproducibility.
Average viable cell yield was 1.7×10(6)/cm(2) of tissue, with 75.5% of the total cell isolate viable. Total viable cell number was not significantly reduced after 4 h storage at 22°C or 4°C, and was stable for 24 h at 4°C. Proliferative potential was assessed using a colony forming assay, with 0.3% of viable cells isolated forming keratinocyte colonies. Predominantly the suspension contained keratinocytes (64.3±28.8%) and fibroblasts (30.3±14.0%), with a small population of melanocytes also identified (3.5±0.5%). Finally, the supernatant contained low total protein (0.92 mg/ml) and the supernatant had no significant effects on cell viability or growth when applied ex vivo.
These results suggest the ReCell(®) device provides a method for the preparation of a cell suspension with high viability and proliferative potential, containing viable melanocytes and no apparent toxic cell debris. Further work on the sustained viability of these cells in vivo, and in particular after application to the wound, will be important to better understand the potential of the ReCell(®) device in the clinic.
使用未经培养的自体细胞来促进伤口愈合和进行重建手术的情况越来越多。一种常见的制备这些细胞的方法是使用 ReCell(®)设备。然而,尽管该设备目前已经在临床上使用,但尚未发表关于使用 ReCell(®)设备制备的细胞悬液的特征描述。
对 ReCell(®)设备应用于伤口的细胞悬液的细胞类型、活力、产量、稳定性和增殖潜能进行特征描述。
使用 ReCell(®)设备从使用切皮刀分离的 2 cm(2)面积的断层皮片中收获细胞。对所得细胞悬液进行细胞产量、细胞类型、随时间的活力、增殖潜能和重现性分析。
平均活细胞产量为 1.7×10(6)/cm(2)组织,总细胞分离物中 75.5%为活细胞。在 22°C 或 4°C 下储存 4 小时后,总活细胞数量没有明显减少,在 4°C 下稳定 24 小时。通过集落形成测定评估增殖潜能,分离的 0.3%活细胞形成角质形成细胞集落。分离的细胞悬液主要包含角质形成细胞(64.3±28.8%)和成纤维细胞(30.3±14.0%),也识别出少量黑素细胞(3.5±0.5%)。最后,上清液中总蛋白含量低(0.92 mg/ml),当在体外用上清液处理时,对细胞活力或生长没有显著影响。
这些结果表明,ReCell(®)设备提供了一种制备高活力和增殖潜能细胞悬液的方法,其中包含有活力的黑素细胞,并且没有明显的有毒细胞碎片。进一步研究这些细胞在体内的持续活力,特别是在应用于伤口后,对于更好地理解 ReCell(®)设备在临床中的潜力非常重要。