I am trying to think of a way to do 3D cell culture with core needle biopsies from patients with Stage 3/Stage 4 lung tumors (the sample is like a 1cm long piece of string). Trypsinizing or treating them enzymatically and growing them in monolayer changes their phenotype quite a bit. I am looking for a method to grow the biopsy samples in vitro, with the goal to keep the phenotype as close as it can be to the in vivo tumor and perform cytotoxicity studies. Any advice or suggestions will be greatly appreciated. Thank you.
Hi
I am also doing 3D cell culture using biopsy samples but not with core needle biopsies. The tissues are enzymatically treated to isolate cells and centrifuge it, the pellet will be seeded on the scaffolds. But I dont know whether u are using a scaffold for 3D cell culture.
Hi Mamatha. Thank you so much. I am not using a scaffold. I am planning on using the hanging drop method initially to form the spheroids and then transferring those to low attachment plates to let them grow bigger, before treating them with drugs. I don't know how to grow the biopsy samples in a way that they don't change their phenotype or differentiate into other cells.
I think it is best way to use hanging plate method but it will give you spheroid but not the in vivo like structure that you wish for. But it is the best method to start with. I can suggest that before going to hanging plate, coat a small layer of agarose of 0.7% up to 500 micron to 1 cm height in 96 well or slides or on cover slip (keep coated cover slip in 6 well plate). The try to cell culture on it and see what kind of 3d structure you are getting under fluorescent or confocal microscope. This is very basic technique you can start with at very low cost. Agarose very much cell friendly. This way you can get much better idea about your sample growth property that will help you further. I hope this answer will help you.
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