Rapid and non-enzymatic in vitro retrieval of tumour cells from surgical specimens

The study of tumourigenesis commonly involves the use of
established cell lines or single cell suspensions of primary
tumours. Standard methods for the generation of short-term tumour
cell cultures include the disintegration of tissue based on
enzymatic and mechanical stress. Here, we describe a simple and
rapid method for the preparation of single cells from primary
carcinomas, which is independent of enzymatic treatment and feeder
cells. Tumour biopsies are processed to 1 mm(3) cubes termed
explants, which are cultured 1-3 days on agarose-coated well plates
in specified medium. Through incisions generated in the explants,
single cells are retrieved and collected from the culture
supernatant and can be used for further analysis including in vitro
and in vivo studies. Collected cells retain tumour-forming capacity
in xenotransplantation assays, mimic the phenotype of the primary
tumour, and facilitate the generation of cell lines.