Abstract
Mesenchymal cells in the crypt play indispensable roles in the maintenance of intestinal epithelial homeostasis through their contribution to the preservation of stem cells. However, the acquisition properties of the production of stem cell niche factors by the mesenchymal cells have not been well elucidated, due to technical limitations regarding the isolation and subsequent molecular and cellular analyses of cryptal mesenchymal cells. To evaluate the function of mesenchymal cells located at the large intestinal crypt, we established a novel method through which cells are harvested according to the histologic layers of mouse colon, and we compared cellular properties between microenvironmental niches, the luminal mucosa and crypts. The gene expression pattern in the cryptal mesenchymal cells showed that receptors of the hormone/cytokine leptin were highly expressed, and we found a decrease in Wnt2b expression under conditions of leptin receptor deficiency, which also induced a delay in cryptal epithelial proliferation. Our novel stratified layer isolation strategies thus revealed new microenvironmental characteristics of colonic mesenchymal cells, including the intrinsic involvement of leptin in the control of mucosal homeostasis.
Highlights
Mesenchymal cells in the crypt play indispensable roles in the maintenance of intestinal epithelial homeostasis through their contribution to the preservation of stem cells
Cryptal mesenchymal cells are considered to be important in mucosal regeneration and homeostasis[18]; we examined whether the leptin system is involved in the production of cryptal niche factors
We evaluated leptin receptors (LepRs)-deficient mice (Leprdb/db) regarding their gene expression levels of representative intestinal stem cell niche factors derived from cryptal CD90+ gp38+ mesenchymal cells, including WNTs, Bone Morphogenetic Proteins (BMP), GREMs, or transcription factors (e.g., Gli[1] and forkhead box l1 (Foxl1)) (Fig. 4b)[23]
Summary
Mesenchymal cells in the crypt play indispensable roles in the maintenance of intestinal epithelial homeostasis through their contribution to the preservation of stem cells. To evaluate the function of mesenchymal cells located at the large intestinal crypt, we established a novel method through which cells are harvested according to the histologic layers of mouse colon, and we compared cellular properties between microenvironmental niches, the luminal mucosa and crypts. Our novel stratified layer isolation strategies revealed new microenvironmental characteristics of colonic mesenchymal cells, including the intrinsic involvement of leptin in the control of mucosal homeostasis. To evaluate the precise function of mesenchymal cells located at the large intestinal crypt, reliable isolation methods of cryptal cells based on the histological layers of the colon are required. We present our novel gut mucosal layer isolation method for the characterization of the mesenchymal cell population in murine colon Using this strategy, we discovered important roles of intrinsic leptin and its involvement in intestinal mucosal regeneration
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