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An In Vitro Model for Studying Cellular Transformation by Kaposi Sarcoma Herpesvirus
Published on: August 25, 2017
PKH26 can transfer to host cells in vitro and vivo
Peng Li1, Run Zhang, Haitao Sun
1Department of Neurosurgery, Southern Medical University, Zhujiang Hospital, Guangzhou, China.
Abstract:
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.
Insights
PKH26 fluorescent dye, commonly used for tracking transplanted cells, is not ideal. Its debris can cause non-labeled cells and tissues to fluoresce, compromising specificity in cell tracing studies.
Area of Science:
- Cell Biology
- Biotechnology
- Stem Cell Research
Background:
- PKH26 fluorescent dye is widely used for cell membrane labeling and tracking transplanted cells.
- Concerns exist regarding the specificity of PKH26, particularly the influence of cell debris from labeled cells.
Purpose of the Study:
- To investigate the impact of cell debris from PKH26-labeled adipose-derived stem cells (ADSCs) on unlabeled cells in vitro and host tissues in vivo.
- To evaluate the specificity of PKH26 as a cell tracer.
Main Methods:
- Assessed PKH26 toxicity and proliferation effects on ADSCs using Cell Count-8 and alamar blue assays.
- Co-cultured PKH26-labeled ADSC debris with unlabeled ADSCs in vitro.
- Injected PKH26-labeled ADSC debris intravenously into rats to assess in vivo effects.
Main Results:
- PKH26 did not inhibit ADSC proliferation or exhibit toxicity compared to unlabeled ADSCs.
- Cell debris from PKH26-labeled ADSCs induced red fluorescence in unlabeled cells in vitro.
- Red fluorescence was observed in host tissues in vivo after injection of PKH26-labeled cell debris.
Conclusions:
- PKH26 is not a specific cell tracer due to fluorescence transfer from its debris.
- The widespread use of PKH26 for cell tracing may lead to inaccurate results.
- Alternative cell tracing methods with higher specificity are needed.
