Abstract
The isolation of viable enterocytes, villi and crypts from the small intestine of a feral bird (Columba livia) is important for performing physiological experiments in ecologically relevant processes of membrane transport. The effectiveness of mechanical disruption, enzymatic digestion and chelating agents were compared. The objectives were to isolate enterocytes, villi and crypts from the small intestine of young pigeons; to evaluate the viability of the isolated intestinal epithelial cells isolated; and to verify the integrity of enterocytes by biochemical features. Enzymatic and mechanical methods yielded both elongated columnar and spherical cells. With the chelating method villi and crypts were obtained. All methods produced a high yield of intestinal epithelial cells with about 50% viability. Brush border enzymes (sucrase-isomaltase and alkaline phosphatase) activities were high and, as reported in chickens, they did not differ along the intestinal villus-crypt axis. Although the three methods have good viabilities, the enzymatic technique gives the best yield in cell number, while the chelating method provides the highest populations of morphologically distinctive villi and crypts.
Highlights
IntroductionThe isolation of intestinal epithelial cells, villi and crypts is important for studies of digestive physiology including developmental physiology (Traber, 1999; Uni et al, 1997; King et al, 2000; Uni et al, 2003), membrane transport processes (Audus et al, 1990; Garriga et al, 1997; Artursson and Borchardt, 1997; Wolffram et al, 1998; Angelo et al, 2002), digestive enzyme activity in different maturation stages of the enterocyte (Traber, 1990; Ferraris et al, 1992; Fan and Stoll, 2001; Uni et al, 1998; Semenza et al, 2001) and cellular biotransformation of agrochemicals (Kurihara et al, 1993)
Most avian intestinal cell studies were performed in chickens (Uni et al, 1998; Angelo et al, 2002; Velge et al, 2002) while we are interested in obtaining intestinal cells from a feral bird (Columba livia) for studies on intestinal membrane transport processes with ecological relevance (Caviedes-Vidal, 2003)
The small intestinal epithelium constitutes a system in constant renewal
Summary
The isolation of intestinal epithelial cells, villi and crypts is important for studies of digestive physiology including developmental physiology (Traber, 1999; Uni et al, 1997; King et al, 2000; Uni et al, 2003), membrane transport processes (Audus et al, 1990; Garriga et al, 1997; Artursson and Borchardt, 1997; Wolffram et al, 1998; Angelo et al, 2002), digestive enzyme activity in different maturation stages of the enterocyte (Traber, 1990; Ferraris et al, 1992; Fan and Stoll, 2001; Uni et al, 1998; Semenza et al, 2001) and cellular biotransformation of agrochemicals (Kurihara et al, 1993). Stem cells found at the base of the crypts continually divide and provide the source of all the epithelial cells in the crypts and on the villi (Booth and O’Shea, 2002)
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