I have genetically modified mouse embryonic stem (ES) cells together with control ES cells. I am planning to inject these into opposing flanks of nude mice. In other words, I will inject the wild-type (WT) ES cells in one flank and the genetically modified cells in the opposite flank. The genetically modified cells overexpress some potential oncogenes and what I want to show is that they will have a higher tumorigenicity, as compared to WT stem cells i.e. perhaps the size and/or malignancy of the tumors will be different.
I have tried to find similar studies (using ES cells injected in mouse flanks to study cancer) but have not been able to find any. Does anyone know of any studies on this? What do you think about it as an experimental strategy?
Stem tumor cells should be implanted uniquely different mice! Accordingly, there should be 2 groups! If a mouse implanted with different sides of the stem and cancer cells, they will affect each other.
Please refer to some of the original work done by Roy Stevens during 60s on various mouse models. Most of the mouse cell lines were the result of his work at The Jackson Laboratories.
http://www.the-scientist.com/?articles.view/articleNo/12717/title/A-Stem-Cell-Legacy--Leroy-Stevens/
the above link has several references. Some of the cell lines are temperature sensitive i.e they differentiate into various lineage . Some of the cell lines are also density sensitive
If you dont want to study teratoma formation,
why would you want to use ES cells as a cancer model?
Thanks a lot for your answers!
Our goal is to use these ES cells to generate transgenic mice. However, this will take a long time and I was thinking whether there is any point in doing any in vivo experiments with these cells right now, like injecting them into mouse flanks. I guess I will have to analyze the formation of teratomas but the main thing is to show whether these genetically modified cells have a higher tumorigenicity and behave differently with respect to proliferation, invasion etc?
Spyridon,
Then you should make transgenic mice (replace one cell in a radiation sterilized 4-cell stage embryo and reimplant in mother) from the control ES cell and then test the results of transplantation into these mice of the control vs modified cells. Otherwise you will have more differences than only the genetic modification. It will probably also be important into which site you do the injections as even normal cells in the wrong location can form teratomas. So you may want to compare flank vs breast fat pad, vs brain, etc.
Hi Spyridon, I agree with the previous comments of Nikita and C. Lange, and would add the some well thought-out pilot studies where you implant cells into different locations (small n of mice suffice) will help you to get the basics correct. Possibly, those cells do not grow at all in the flank (will you add matrigel? will you inject intradermal or really subcutaneous, which is a difference?). SOme time ago, wedid such pilot studies with hESC which was published, see
https://www.researchgate.net/publication/24359332_Teratoma_formation_by_human_embryonic_stem_cells_evaluation_of_essential_parameters_for_future_safety_studies?ev=prf_pub
All the best for your studies Hannes
Article Teratoma formation by human embryonic stem cells: Evaluation...
- Badges
- Science topic