A metastatic process is a 3D process occuring in an amazingly complex tumor microenvironment.

Any cell migration / invasion assays relating to "pure cancer cells forced to evolve in 2D dimension and prisoners of closed plastic dishes" have nothing to do with the actual 3D journey of metastatic cancer cells.

Here attached are various in vitro approaches that recapitulate some of the important aspects of an actual metastatic process.

Best regards

PS: I also added 3 reviews for those of you who would be interested by the metastatic journey. These are the first three articles here attached.

There are quite a number of in vitro assays to determine the metastatic potential of cells but I would suggest the most widely and accepted Cell Migration assay....a lot of companies sell kits based on this assay and it is widely accepted.All the best.

Interesting question. Metastatic potential measured by cell migration assays in vitro would be more associated with local invasion and migration of tumour cells rather than metastatic colonization or distant organ-specific colonization which is seen in some metastatic tumours.

In the case of migration assays, you don't know if you model will metastasize or rather grow diffusely or invasively in vivo until you actually do the experiment.

You might think about boyden chamber assay, gap closure assay (scratch assay) and anchorage independent growth assays in soft agar. All of them will give just hints, the in vivo experiment has to be done as well.

Thanks everyone for the input. I already did scratch assay and soft agar assay. I found positive result. But problem is that..Epithelial-mesenchymal transition (EMT) and MET plasticity is crucial for efficient metastatic colonization. So, even though my target gene prevents migration and induces MET, I can`t be sure whether this is for metastasis suppression or metastatic colonization.

A metastatic process is a 3D process occuring in an amazingly complex tumor microenvironment.

Any cell migration / invasion assays relating to "pure cancer cells forced to evolve in 2D dimension and prisoners of closed plastic dishes" have nothing to do with the actual 3D journey of metastatic cancer cells.

Here attached are various in vitro approaches that recapitulate some of the important aspects of an actual metastatic process.

Best regards

PS: I also added 3 reviews for those of you who would be interested by the metastatic journey. These are the first three articles here attached.

Thank you so much Robert for the input.

I recently did the invasion (using pre-matrigel coated invasion insert) and migration assay (insert without matrigel) of TNBC cells under a particular gene control. I found great finding that is consistent with in vivo mouse model. If you want to discuss on it I can provide you the related information and possible trouble shoot that you may encounter. Optimization of time period is also crucial as well. Good luck.

Thank you Iqbal. Actually this question was raised by me 3 year ago while I was a graduate student. I did invasion (in matrigel coated insert) and migration assay a lot at that time and did not have any problem. Metastatic colonization, which is a very complex "IN VIVO" process and its really hard to demonstrate in vitro. To some extent, it has nothing to do with in vitro invasion/migration assay as there is no stroma and immune involvement. Let me know if you need more info about metastatic colonization.