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Abstract
Understanding the nature of cell surface markers on exfoliated colonic cells is a crucial step in establishing criteria for a normally functioning mucosa. We have found that colonic cells isolated from stool samples (SCSR-010 Fecal Cell Isolation Kit, NonInvasive Technologies, Elkridge, MD), preserved at room temperature for up to one week, with viability of >85% and low levels of apoptosis (8% - 10%) exhibit two distinct cell size subpopulations, in the 2.5 μM - 5.0 μM and 5.0 μM - 8.0 μM range. In addition to IgA, about 60% of the cells expressed a novel heterodimeric IgA/IgG immunoglobulin that conferred a broad-spectrum cell mediated cytotoxicity against tumor cells. In a cohort of 58 subjects the exclusive absence of this immunoglobulin in two African-Americans was suggestive of a germline deletion. Serial cultures in stem cell medium retained the expression of this heterodimer. Since a majority of the cystic cells expressed the stem cell markers Lgr5 and Musashi-1 we termed these cells as gastrointestinal progenitor stem cells (GIP-C**). CXCR-4, the cytokine co-receptor for HIV was markedly expressed. These cells also expressed CD20, IgA, IgG, CD45, and COX-2. We assume that they originated from mature columnar epithelium by dedifferentiation. Our observations indicate that we have a robust noninvasive method to study mucosal pathophysiology and a direct method to create a database for applications in regenerative medicine.
Highlights
The human colon renews its mucosa almost every five days and allows the colonization of the lumen by a host of symbionts, microorganisms referred to collectively as the colonic microbiome
In this report we propose a sentinel mechanism involving the dedifferentiation of exfoliated colon epithelial cells to cystic progenitor stem cells (GIP-C) expressing the intestinal transit stem cell markers Lgr-5 and Musashi-1 (Msi 1)
About 10% of the exfoliated colonic cells undergo apoptosis—Viability of exfoliated colonic cells isolated from fecal samples (“somatic cell sampling and recovery” (SCSR) Cells”) was assessed by trypan blue exclusion (85%), an observation that was confirmed by flow cytometry using propidium iodide exclusion
Summary
The human colon renews its mucosa almost every five days and allows the colonization of the lumen by a host of symbionts, microorganisms referred to collectively as the colonic microbiome This rapid and constant renewal of the mucosa requires a steady source of proliferating cells to maintain a stable turnover rate. Cells lining the colonic mucosa acquire the ability to tolerate foreign antigens generated by commensal microflora along with a mixture of antigens of dietary origin. This is made possible through a process of immunological acquiescence resulting in a dynamic equilibrium, which requires the cells of the mucosa to exhibit an adaptive tolerance to the microflora unique to each individual. Tropical sprue is a syndrome associated with substantial colonization of the proximal small bowel with coliform bacteria that are normally harmless when present in the large bowel [7]
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