Dissection of tumour and host cells from target organs of metastasis for testing gene expression directly ex vivo.
We report on a new methodology which allows the direct analysis ex vivo of tumour cells and host cells (lymphocytes, macrophages, endothelial cells) from a metastasised organ (liver or spleen) at any time point during the metastatic process and without any further in vitro culture. First, we used a tumour cell line transduced with the bacterial gene lacZ, which permits the detection of the procaryotic enzyme beta-galactosidase in eukaryotic cells at the single cell level thus allowing flow adhesion cell sorting (FACS) analysis of tumour cells from metastasised target organs. Second, we established a method for the separation and enrichment of tumour and host cells from target organs of metastasis with a high viability and reproducibility. As exemplified with the murine lymphoma ESb, this new methodology permits the study of molecules of importance for metastasis or anti-tumour immunity (adhesion, costimulatory and cytotoxic molecules, cytokines, etc.) at the RNA or protein level in tumour and host cells during the whole process of metastasis. This novel approach may open new possibilities of developing strategies for intervention in tumour progression, since it allows the determination of the optimal window in time for successful treatments. The possibility of direct analysis of tumour and host cell properties also provides a new method for the evaluation of the effects of immunisation with tumour vaccines or of gene therapy.
Rocha
M
M
Tumor Immunology Program, German Cancer Research Center, Heidelberg, Germany.
Hexel
K
K
Bucur
M
M
Schirrmacher
V
V
Umansky
V
V
D000818:Animals; D002450:Cell Communication; D004210:Dissection; D004727:Endothelium / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression; D007728:Kupffer Cells / Q000166:cytology / Q000378:metabolism; D007763:Lac Operon; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000378:metabolism* / Q000473:pathology / Q000556:secondary*; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism; D008223:Lymphoma / Q000378:metabolism* / Q000473:pathology*; D008264:Macrophages / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008811:Mice, Inbred DBA; D013154:Spleen / Q000166:cytology* / Q000378:metabolism; D013160:Splenic Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000556:secondary*; D014161:Transduction, Genetic