Dear Duje,

The response to your question highly relates to the design of your experiment and to the cancer cell type you are using.

You are not obliged to go to s.c. inoculation: you can go on other sites, but once more it is dependent on the type of experiment you want to perform. You can go on i.p. route for example. You can go on various orthotopic transplantation, etc..

You can also put your "3D structures" into specific "ad hoc" chambers and implant them s.c.

etc...

Best regards

Robert 

The idea is to produce a spheroid from a549 and a549-epoB40 cell line trom lung cancer, to implant it into the mouse and look if They act the same as in culture. A549 are usually implanted s.c. in xenografts model, but as single cells large number is needed. So I am a bit concerned if quite small steroid would survive implantation.

thank you.

You can indeed use A549 NSCLC cells s.c. in nude mice (see the attached article).

However, it is not a "s.c. NSCLC" that kills the patient, but the metastases from the primary lung tumor.

The A549 NSCLC in orthotopic xenograft mimic well what happens in clinics (see Mathieu et al., 2004).

Thus, if you want to see the differences between 2D and 3D cultured A549 cells when implanted in vivo, you actually should go on orthotopic xenografts, not s.c. ones.

The differences between 2D and 3D in vitro cultures will increase the metastatic abilities of "3D cultured A549 cells", a feature that you will not be able to see with s.c. grafting.

I am quite sure that with a low number of 3D A549 cells, you will have positive tumorigenesis in orthotopic implantation, but not in s.c. one.

You can even go with intra-cerebral grafting of A549 cells (see Ingrassia et al), a procedure that will allow you to further decrease (as compared to lung orthotopic) the number of cells needed for in vivo tumorigenesis.

Brain metastases is frequent for NSCLCs, and you can thus use a "NSCLC pseudo-metastatic model".

Best regards

Robert