Brief report: CD24 and CD44 mark human intestinal epithelial cell populations with characteristics of active and facultative stem cells

Abstract

Recent seminal studies have rapidly advanced the understanding of intestinal epithelial stem cell (IESC) biology in murine models. However, the lack of techniques suitable for isolation and subsequent downstream analysis of IESCs from human tissue has hindered the application of these findings toward the development of novel diagnostics and therapies with direct clinical relevance. This study demonstrates that the cluster of differentiation genes CD24 and CD44 are differentially expressed across LGR5 positive “active” stem cells as well as HOPX positive “facultative” stem cells. Fluorescence-activated cell sorting enables differential enrichment of LGR5 cells (CD24−/CD44+) and HOPX (CD24+/CD44+) cells for gene expression analysis and culture. These findings provide the fundamental methodology and basic cell surface signature necessary for isolating and studying intestinal stem cell populations in human physiology and disease.

Orphan Gpr182 suppresses ERK-mediated intestinal proliferation during regeneration and adenoma formation

“…+ cells, as previously reported (40,41). Neither the percentage of CD326 + epithelial cells nor the proportion of CD44 -and CD44 + epithelial cells was altered with the loss of Gpr182 (Supplemental Figure 6, A and B Figure 5H).…”

“…Single-cell isolation and ex vivo culture. Single cells from jejunum of 8-week-old female Gpr182 +/+ CMV-Cre and Gpr182 Δ/Δ CMV-Cre mice were isolated and cultured as previously described (31,32,(40)(41)(42)64). Briefly, jejuna were flushed and incubated in 3 mM EDTA (Corning) and 10 μM Y27632 (Selleck Chemicals) in Dulbecco's PBS.…”

“…Tissues were either snap-frozen in liquid nitrogen or stored in RNAlater (Ambion, Thermo Fisher Scientific). Enriched cell populations labeled with CD326 and CD44 or differential Sox9-EGFP or Lgr5-EGFP levels as previously described (18,30,(40)(41)(42) were isolated by FACS and collected into RNAqueous-Micro Lysis Solution (Ambion, Thermo Fisher Scientific). Likewise, the Paneth cell-enriched population was isolated using a previously published protocol (31).…”

Utility of models of the gastrointestinal tract for assessment of the digestion and absorption of engineered nanomaterials released from food matrices

“…Simple cell culture systems using established cell lines do not incorporate factors such as age and sex, and they lack systemic circulation pathways such as the enterohepatic secretion of absorbed materials back into the GI lumen. Emerging approaches to account for different genotypes could possibly be developed, such as the use of induced pluripotent stem cells (Gracz et al, 2013). The InLiveTox project (http://www.inlivetox.eu/) has developed a multi-tissue fluidic model that allows flow above and below a Caco2 differentiated confluent layer, with the basolateral flow linking to endothelial (HUVEC) and hepatocyte (C3A) culture.…”

Intestinal stem cells and stem cell-based therapy for intestinal diseases

“…Moreover, epherin receptors (Ephs) mediate cell-cell communications and were reported to be ISC markers expressed at the crypt base . Recently, CD24 and CD44 could be used to FACSisolate human intestinal epithelial cell populations with characteristics of active and facultative stem cells (Gracz et al 2013). A similar study introduced CD24 as a membrane marker used for the isolation of mouse ISC fraction with the characteristics of stemness.…”