ANALYSIS OF TUMOR-SPECIFIC ALTERATIONS IN NATIVE SPECIMENS BY PCR - HOW TO PROCURE THE TUMOR-CELLS (REVIEW)

Native tumours, in contrast to cell lines, are usually heterogeneous, consisting of tumour cells, stroma, infiltrating leukocytes, necrotic cells, and surrounding normal tissue. Therefore, when using non-linear amplification and detection methods such as the PCR, the verification of the DNA from the tumour cells is mandatory to avoid equivocal or f alse results. Here, current methods to isolate tumour cells from nativ e tumours are reviewed. The methods are: i) a variety of microdissecti on techniques including microdissection of membrane-mounted native tis sue (MOMeNT), ii) Selective ultraviolet radiation fractionation (SURF) , iii) antibody-based tumour cell selection and flow cytometric cell o r cell nucleus sorting, and iv) in situ PCR. Each of the methods has b een used, and overall preference cannot be given to any of them. Accur acy, reproducibility, documentation, cost, and applicability in a rout ine setting are discussed, from which fields of preferencial use may e merge for the different methods.