Hi All,
A friend of mine from India has raised a concern since they don't have the transduction facility, what are the other ways to trace the transplanted Stem Cells in vivo in rodents if they are not virally transduce for any color e.g. GFP/mCherry etc. Their lab is planing to transplant the mouse NSCs in mouse only.
Kindly please suggest.
Many thanks,
Syed
If you transplant male cells into a female recipient, they can be detected by FISH using a Y chromosome probe.
One option could be use male derived stem cells in female recipient and then do fish to locate Y chromosome in sample. Any cell with Y chromosome then should be derived from transplanted cells (at least in virgin mouse).
I have had a customer successfully use DiR to fluorescently label and track stem cells in vivo, and they can be visualized without sacrificing the animal...
Could your isolate NSCs from transgenic mice that are able to express GFP and luciferase? The luciferase would allow for long term in vivo monitoring (bioluminescence imaging). In this case you don't need to transfect or transduce the cells, but at the same time you are sure that the signal doesn't get diluted with cell proliferation.
If you transplant male cells into a female recipient, they can be detected by FISH using a Y chromosome probe.
I aggree with JJ Limnios. Implanted stem cells from a male donor, i;e; having aY chromosone, can always be recognized among cels of thez female, i.e. XX recipient. It is necessary to have the Y probe ... Good luck
the only way is using male NSCS to female mice and then looking for Y chromosome by FISH
I would second the use of CFSE - even at low concentrations it is very bright and the stain is relatively long lived. I would imagine that over the course of a few weeks you should still be able to reliably see the dye.
Hi Syed
an easy method could be hoechst labelling of nuclei (see in example "Longitudinal Tracking of Human Fetal Cells Labeled with Super Paramagnetic Iron Oxide Nanoparticles in the Brain of Mice with Motor Neuron Disease" by Bigini et al, Plos one Published: February 27, 2012, DOI: 10.1371/journal.pone.0032326).
Anyway you may have some problems with dye diffusion so it' a perfect tool for preliminary screening but need additional support/tecnique to garantee pubblication.
Hope it could be useful...good luck!
It is possible to purchase EGFP mice from Jackson Laboratory http://jaxmice.jax.org/strain/003291.html. These mice prepared by Dr. Okabe have excellent fluorescence in neural tissue and in muscles. We have transplanted CAG-GFP stem cells to C57Bl/6 congenic recipients and easily identified their progeny in brain by microscope. If you want to observe accumulation of NCSs in certain tissue (let's say tumor) in vivo, you must use preclinical optical imager. The results depend on sensitivity of the instrument (there are many on the market) and on number of cells/strenght of the signal. Another possibility is to mark cells with SPIOs and observe them on microMRI or MPI but I don't expect that they have access to these expensive technologies. Similar situation is with radioisotope labeling for PET or SPECT.
- Badges
- Science topic
- Similar topics
- Methodology
- Laboratory
- Cellular Biology Techniques