Human colon tissue in organ culture: preservation of normal and neoplastic characteristics.
Normal and neoplastic human colon tissue obtained at surgery was used to establish conditions for organ culture. Optimal conditions included an atmosphere of 5% CO2 and 95% O2; tissue partially submerged with mucosa at the gas interface; and serum-free medium with 1.5 mM Ca2+ and a number of growth supplements. Histological, histochemical, and immunohistochemical features that distinguish normal and neoplastic tissue were preserved over a 2-d period. With normal tissue, this included the presence of elongated crypts with small, densely packed cells at the crypt base and mucin-containing goblet cells in the upper portion. Ki67 staining, for proliferating cells, was confined to the lower third of the crypt, while expression of extracellular calcium-sensing receptor was seen in the upper third and surface epithelium. E-cadherin and β-catenin were expressed throughout the epithelium and confined to the cell surface. In tumor tissue, the same disorganized, abnormal glandular structures seen at time zero were present after 2 d. The majority of cells in these structures were mucin-poor, but occasional goblet cells were seen and mucin staining was present. Ki67 staining was seen throughout the abnormal epithelium and calcium-sensing receptor expression was weak and variable. E-cadherin was seen at the cell surface (similar to normal tissue), but in some places, there was diffuse cytoplasmic staining. Finally, intense cytoplasmic and nuclear β-catenin staining was observed in cultured neoplastic tissue.
Dame
Michael K
MK
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI 48109, USA. mdame@med.umich.edu
Bhagavathula
Narasimharao
N
Mankey
Cohra
C
DaSilva
Marissa
M
Paruchuri
Tejaswi
T
Aslam
Muhammad Nadeem
MN
Varani
James
J
D015415:Biomarkers / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002245:Carbon Dioxide / Q000494:pharmacology; D002454:Cell Differentiation; D003106:Colon / Q000254:growth & development / Q000473:pathology*; D003110:Colonic Neoplasms / Q000473:pathology*; D004801:Eosine Yellowish-(YS) / Q000378:metabolism; D006416:Hematoxylin / Q000378:metabolism; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D009924:Organ Culture Techniques; D010100:Oxygen / Q000494:pharmacology; D044169:Receptors, Calcium-Sensing / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
D015415:Biomarkers; C095550:CASR protein, human; D015820:Cadherins; D044169:Receptors, Calcium-Sensing; D051176:beta Catenin; D002245:Carbon Dioxide; D020781:Matrix Metalloproteinase 1; D010100:Oxygen; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin