Isolation of the Epithelial Cells of the Rat Small Intestine

Perris, A. D.

doi:10.1139/o66-086

Cited by 57 publications

(27 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The methods that have been developed for the isolation of columnar epithelial cells (enterocytes) from the small intestine of the rat have been applied to the adult animal [Harrison and Webster, 1969;Huang, 1965;Kimmich, 1970;Perris, 1966;Sjostrand, 1968;Towler et al, 1978]. There have been no studies comparing the function of cells isolated from the jejunum to those isolated from the ileal region nor the relative ability of all of the various methods for isolating metabolically intact cells.…”

Section: Discussionmentioning

confidence: 99%

Comparison of the Metabolic Activities of Enterocytes Isolated from Different Regions of the Small Intestine of the Neonate

Nm¹

1985

Neonatology

View full text Add to dashboard Cite

Enterocytes were isolated from the jejunum and ileum of the neonatal (12-day-old) rat by vibrating everted intestinal segments in buffered saline containing EDTA. The isolated cells appeared morphologically normal when examined microscopically, retaining their in vivo shape and internal structure. Enzyme analysis revealed that over 80% of the total gut lactase activity was associated with the isolated enterocytes. During incubation at 37 °C the cells maintained their structure for 30–60 min and consumed oxygen and produced lactate at a constant rate for at least 45 min. On the basis of these tests it was concluded that the method yielded large numbers of enterocytes which were able to be maintained in simple media without significant loss of structure or function for at least 45 min at 37 °C. Comparison of the metabolic activities of cells isolated from the two regions suggests that ileal enterocytes maintain the greater functional integrity, maintaining an intact plasma membrane for longer periods and having higher respiratory and glycolytic activities.

show abstract

Section: Discussionmentioning

confidence: 99%

Comparison of the Metabolic Activities of Enterocytes Isolated from Different Regions of the Small Intestine of the Neonate

Nm¹

1985

Neonatology

View full text Add to dashboard Cite

show abstract

“…The volume of the chamber was about 0.1 ml. All solutions for the isolation or storage of the cells except those used for patching contained 0.1% bovine serum albumin, which is known to have a "protective" effect on isolated enterocytes (Perris, 1965). During isolation, most of the enterocytes rounded off and attached to the glass bottom of the chamber.…”

Section: Cell Isolationmentioning

confidence: 99%

Single chloride channels in colon mucosa and isolated colonic enterocytes of the rat

et al. 1989

View full text Add to dashboard Cite

Chloride channels from rat colonic enterocytes were studied using the patch-clamp technique. After removal of mucus, inside-out patches were excised from the apical membrane of intact epithelium located at the luminal surface. They contained spontaneously switching Cl- channels with a conductance of 35-40 pS. The channels were blocked reversibly by anthracene-9-carboxylic acid (1mM). In excised patches from single enterocytes, isolated by calcium removal, the Cl- channels were studied in more detail. The I-V relation was linear between +/- 80 mV. The selectivity was I- greater than Br- greater than Cl- = NO3- greater than F- = HCO3-. Thirty pS Cl- channels were also found on the basolateral membrane of crypts isolated by brief calcium removal. The I-V curve of these Cl- channels was also linear. The results provide direct evidence for the existence of Cl- channels in the apical membrane of surface cells in colonic mucosa. The properties of these channels are similar to those previously observed when incorporating membrane vesicles into planar lipid bilayers. Both results support the validity of the theoretical models describing intestinal secretion.

show abstract

“…It would then be possible to avoid the assumption that measurements made on mucosal scrapings necessarily represent the activity of the mucosal epithelium which may in fact constitute only a relatively small and variable proportion of the starting material . Various workers have isolated intestinal epithelial cells into suspension (1)(2)(3)(4)(5)(6)(7), but have made little or no attempt to show that the cells were functionally intact . With any method of tissue disaggregation there is the possibility of cell damage, so it is essential to demonstrate that isolated cells to be used for metabolic studies have retained their structure and function .…”

Section: Introductionmentioning

confidence: 99%

Studies on Epithelial Cells Isolated From Guinea Pig Small Intestine

Evans

Wrigglesworth

Burdett

et al. 1971

The Journal of Cell Biology

View full text Add to dashboard Cite

Sheets of mucosal epithelial cells were released from guinea pig small intestine after incubation with ethylenediaminetetraacetate . Cells in sheets retained their columnar shape for 24 hr at room temperature, and exclusion of nigrosine suggested they had intact plasma membranes . When sheets were disaggregated individual cells had normal morphology for at least 4 hr. During isolation 16% of the total protein and 24% of the total lactic dehydrogenase were lost from the cells, but subsequent enzyme leakage was low . Leakage increased with shaking, incubation at 37°C, or increasing the oxygen tension of the suspending medium, but was minimal when the Na+ : K+ ratio in the medium was 8 :1 and the osmolarity was high . Losses of particulate enzyme activities were negligible . Respiration was constant for up to 4 hr and was insensitive to calcium, bicarbonate, oxygen tension, and pH . It was inhibited by cyanide and iodoacetate and varied with the Na+ : K+ ratio of the extracellular fluid and the structural integrity of the cells . All preparations concentrated potassium and excluded sodium, but lost this ability if ouabain was added or cells were broken . Potassium-42 uptake was also sensitive to temperature, ouabain, and structural integrity. The preparations are being used to study cell metabolism in the intestinal epithelium.

show abstract

Isolation of the Epithelial Cells of the Rat Small Intestine

Cited by 57 publications

References 0 publications

Comparison of the Metabolic Activities of Enterocytes Isolated from Different Regions of the Small Intestine of the Neonate

Comparison of the Metabolic Activities of Enterocytes Isolated from Different Regions of the Small Intestine of the Neonate

Single chloride channels in colon mucosa and isolated colonic enterocytes of the rat

Studies on Epithelial Cells Isolated From Guinea Pig Small Intestine

Contact Info

Product

Resources

About