I would like to track cells after transplantation into the brain with PKH26 .I have several questions regarding to use of this dye:
1 .Does cellular stain with PKH26 last up to two months after cells transplantation ?
2. Which one is better for tracking cells stained with PKH26, frozen section or paraffin block method?
3. Dose perfusion with paraformaldehyde effect on density of stained cells?
4. After removing the brain and during sectioning is it necessary to store the samples in darkness according to the fluorescent property of this dye?
Thank you in advance for your help
Hy,
attached you find an article where I describe the possbility to co-label transplanted cells in the brain. Paraffin embedding is an absolute No-Go for PKH26!!!!
However, 16 years later I would not recommend to use PKH26 for such kind of studies. If you transplant human cells, an antibody against human cell nuclei is the better way to characterize transplanted cells.
Good luck
Stefan
Hi,
If needed, you can stain and use PKH26 stained cells for transplantation in the brain. Normally, the staining will stay for 100 days even if after each cell division, a part of the staining is going with the newly formed cell.
For tracking cells, frozen section is definitely the only way to do.
Normally, PFA will not affect the brain. I did with PKH67 and it was working perfectly so I can assume that il will be the same with PHK26.
For a fluorescent dye, it's always better to work in the dark
Best regards,
Thomas
We used PKH26 for cell tracking after transplantation several years ago, however, the dye leaked out (or perhaps the cells were not washed properly, these days I can't say) and we found several obviously host cells labeled with the dye as well. It was not reliable. For human cells HuNu or MTC02 is much better. For same species (rat to rat) carboxy-fluorescein diacetate succinimidyl ester (CFDA-SE) lipophilic dye worked better than pKH26.
Hi,
I used PKH26 in order to track cells after transplantation in the brain. I could see fluorescence after 3 months. I perfused mice and I had no problem. However, if you have the possibility, I would recommend to use fluorescent cells, such as GFP-expressing cells.
Hello Maryam,
can you please let us know what kind of cell labelling you will now use for your planned experiments?
Sincerely Yours
Stefan
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