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[Grid electron microscopy studies of isolated intestinal epithelium]
Abstract:
Four media are used to isolate enterocytes from the jejunum of the mouse (basic medium with either hyaluronidase, dispase, pronase, or EDTA). During incubation the intestinal wall is gently agitated. Layers of epithelial cells rather than single cells are removed from the mucosal surface by these procedures. Small groups of enterocytes and single cells can be obtained by further mechanical agitation, e.g. by repeatedly washing the isolated epithelia. Similar morphological findings (transmission and scanning electron microscopy) are obtained after using the various isolation media. Only with EDTA a dilatation of the endoplasmic reticulum is regularly seen. Isolation of the enterocytes starts from the tip of the intestinal villus. The straight apical surface becomes convex and the most lateral microvilli of the brush border desintegrate. The opening of the terminal bars can be observed. Surface differentiations on the lateral cell surfaces of isolated enterocytes become less obvious and disappear. This is discussed with respect to the so-called "lateral vacuoles" which appear in isolated enterocytes. Large spherical protrusions are developed at the basal surface of enterocytes. The cell membrane covering these protrusions is frequently found to be discontinuous. No other cell organelles than ribosomes are to be seen inside of these protrusions. Membrane-bound vesicles are also frequently seen. It is suggested that the basal spherical protrusions are casted off.
Insights
This study details methods for isolating mouse jejunal enterocytes using various media. Researchers observed morphological changes in isolated cells, including basal protrusions, offering insights into cell structure and isolation techniques.
Area of Science:
- Cell Biology
- Gastroenterology
Context:
- Enterocytes, the primary cells of the intestinal lining, play crucial roles in nutrient absorption and barrier function.
- Isolation of viable enterocytes is essential for studying their physiology and response to various stimuli.
Purpose:
- To evaluate four different media (hyaluronidase, dispase, pronase, and EDTA) for their efficacy in isolating enterocytes from mouse jejunum.
- To characterize the morphological changes in enterocytes following isolation using transmission and scanning electron microscopy.
Summary:
- Four media were tested for isolating jejunal enterocytes from mice, with gentle agitation yielding epithelial layers.
- Further mechanical agitation produced single cells and small groups.
- Electron microscopy revealed similar morphologies across media, except for EDTA-induced endoplasmic reticulum dilation.
- Observed changes included apical surface alterations, loss of lateral surface differentiations, appearance of lateral vacuoles, and formation of basal spherical protrusions, possibly indicating cell shedding.
Impact:
- Provides a comparative analysis of enterocyte isolation techniques, aiding researchers in selecting optimal methods.
- Detailed morphological characterization of isolated enterocytes offers valuable data for cell biology and gastroenterology research.
- The findings contribute to understanding the structural integrity and potential shedding mechanisms of intestinal epithelial cells post-isolation.