Culture of primary epithelial adenoma cells from familial adenomatous polyposis patients.
BACKGROUND
Colorectal tumors arise from unregulated cell proliferation of the intestinal epithelium through a multistep process the first step usually being premalignant adenomas. Familial adenomatous polyposis patients carry a germ line mutation in the APC gene leading to the development of thousands of polyps, which, if left untreated, lead to cancer. The goal of this study was the establishment of conditions for the culture of epithelial cells composing an adenomatic structure.


METHODS
All colorectal specimens were obtained from FAP patients within 1-2 hours of surgery. Cells were cultured by standard procedures. Characterization was carried out by immunostaining with pancytokeratin, vimentin and desmoplakin antibodies.


RESULTS
A culture protocol that gave rise to epithelial cell growth with high efficiency and efficacy was established. Successful subculturing of the cell sheets took place only when dispase prepared in Ca2+ and Mg2+ free medium, was used to digest polyp tissue taken from FAP patients. By using immunostaining these cells were characterized as epithelial.


CONCLUSIONS
The protocol we developed here provides a means of preparing cell cultures from human colorectal adenomas, which aid in the research of the transition from adenoma to carcinoma.
Molecular Diagnostics Lab, I/RRP, National Center for Scientific Research Demokritos, Athens, Greece.
Florentia
F
Fostira
Angela
A
Apessos
Eutyxia
E
Oikonomou
Panos
P
Kouklis
Sotiris
S
Baratsis
Grigoris
G
Manifikos
Ladislav
L
Andera
Drakoulis
D
Yannoukakos
Alexander
A
Pintzas
George
G
Nasioulas
D000236:Adenoma / Q000473:pathology*; D011125:Adenomatous Polyposis Coli / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods*; D003111:Colonic Polyps / Q000473:pathology; D015179:Colorectal Neoplasms / Q000473:pathology; D004847:Epithelial Cells / Q000473:pathology*; D006801:Humans; D010641:Phenotype