Isolation of exfoliated colonic epithelial cells, a novel, non-invasive approach to the study of cellular markers.
Human stool is a heterogeneous mixture of non-digestible food residues, bacteria, cells exfoliated from the gastrointestinal mucosa and other secretory products. We have demonstrated that fresh human stools dispersed in a buffered saline solution can be fractionated over Percoll/BSA gradients to yield 9 discrete bands of cells in the density range of rho 1.033 to 1.139 and which could be further purified over Histopaque 1077. Enzyme-linked immunoassays (ELISA) for colon-specific antigen (CSA) and cytokeratins (CK) were positive. Western blot analysis showed the presence of 3 cytokeratin bands in the 40-kDa to 60-kDa range suggestive of cytokeratins 8, 18, and 19. Fluorescence flow-cytometric analysis of these cells using antibodies against CSA, CK, the blood-group antigens, carcinoembryonic antigen (CEA), non-mucus-secreting columnar-epithelium-specific MAb PR1A3, and to mucus-secreting colonic-epithelium-specific MAb PR5D5 showed varying degrees of reactivity. Expression of the blood-group phenotype suggests that cells from the proximal half of the colon had survived the transit, since in the adult expression of this marker is limited to cells from the proximal region of the colon. In this report we demonstrate the feasibility of studying, non-invasively, cell-specific markers on exfoliated cells isolated from stools. The evidence strongly suggests that almost all the cells are of colonic origin.
Albaugh
G P
GP
Lipid Nutrition Laboratory, USDA, Beltsville, MD 20705.
Iyengar
V
V
Lohani
A
A
Malayeri
M
M
Bala
S
S
Nair
P P
PP
D000951:Antigens, Neoplasm / Q000032:analysis; D002272:Carcinoembryonic Antigen / Q000032:analysis; D002458:Cell Fractionation; D002469:Cell Separation / Q000379:methods*; D003106:Colon / Q000166:cytology*; D004847:Epithelial Cells; D005243:Feces / Q000166:cytology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007633:Keratins / Q000032:analysis; D008297:Male; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; C030139:colon-specific antigen; D012710:Serum Albumin, Bovine; D007633:Keratins