Department of Neurosurgery, Southern Medical University, Zhujiang Hospital, Guangzhou, China.
Stem Cells Dev. 2013 Jan 15;22(2):340-4. doi: 10.1089/scd.2012.0357. Epub 2012 Oct 19.
The fluorescent dye, PKH26, which mainly binds to the cell membrane, has been used as the cell tracer to locate the transplanted cells in host for a long time. However, there was no detailed report that whether the PKH26 dye was specific to the transplanted cells. Therefore, the aim of this article is to explore the effect of cells debris as the cracking cells from the PKH26-labeled adipose-derived stem cells (ADSCs) on the cells in vitro and the host in vivo. After we tested the proliferation and toxicity of PKH26 to the ADSCs by the Cell Count-8 kit and alamar blue assay, we constructed 2 models, coculturing lots of PKH26-labeled cell debris with the unlabeled ADSCs in vitro and injecting via the tail vein in rat, to evaluate the specificity of the PKH26 dye. The result indicated that the PKH26 didn't inhibit the proliferation and had no toxicity to the ADSCs compared with the unlabeled ADSCs, but the cell debris cracking from PKH26-labeled transplanted cells can cause the unlabeled cells to emit red fluorescence in vitro and also lead the tissues displaying red fluorescence in vivo. We can conclude that the PKH26 dye, used as a cell tracer for a long time, was not an ideal cell tracer.
荧光染料 PKH26 主要与细胞膜结合，长期以来一直被用作示踪移植细胞在宿主中的位置的细胞示踪剂。然而，并没有详细的报告表明 PKH26 染料是否专门针对移植细胞。因此，本文的目的是探讨细胞碎片（从 PKH26 标记的脂肪来源干细胞（ADSCs）中裂解的细胞）对体外细胞和体内宿主的影响。在使用 Cell Count-8 试剂盒和 alamar blue 测定法测试 PKH26 对 ADSCs 的增殖和毒性后，我们构建了 2 种模型，即体外大量共培养 PKH26 标记的细胞碎片和未标记的 ADSCs，以及通过尾静脉注射到大鼠体内，以评估 PKH26 染料的特异性。结果表明，与未标记的 ADSCs 相比，PKH26 不会抑制 ADSCs 的增殖，也没有毒性，但从 PKH26 标记的移植细胞中裂解的细胞碎片可导致体外未标记的细胞发出红色荧光，并且也导致体内组织显示红色荧光。我们可以得出结论，长期以来一直被用作细胞示踪剂的 PKH26 染料并不是一种理想的细胞示踪剂。
