PMID- 29433868 OWN - NLM STAT- MEDLINE DCOM- 20190307 LR - 20190307 IS - 1095-8673 (Electronic) IS - 0022-4804 (Print) IS - 0022-4804 (Linking) VI - 223 DP - 2018 Mar TI - Critical intestinal cells originate from the host in enteroid-derived tissue-engineered intestine. PG - 155-164 LID - S0022-4804(17)30706-0 [pii] LID - 10.1016/j.jss.2017.11.015 [doi] AB - BACKGROUND: Enteroid-derived tissue-engineered intestine (TEI) contains intestinal subepithelial myofibroblasts (ISEMFs) and smooth muscle cells (SMCs). However, these cell types are not present in the donor enteroids. We sought to determine the origin of these cell types and to quantify their importance in TEI development. MATERIALS AND METHODS: Crypts from pan-EGFP or LGR5-EGFP mice were used for enteroid culture and subsequent implantation for the production of TEI. TEI from pan-EGFP enteroids was labeled for smooth muscle alpha actin (SMA) to identify ISEMFs and SMCs and green fluorescent protein (GFP) to identify cells from pan-EGFP enteroids. Fluorescence in situ hybridization (FISH) for the Y chromosome was applied to TEI from male LGR5-EGFP enteroids implanted into female nonobese diabetic/severe combined immunodeficiency mice. To identify chemotactic effects of intestinal epithelium on ISEMFs, a Boyden chamber assay was performed. RESULTS: Immunofluorescence of TEI from pan-EGFP enteroids revealed GFP-positive epithelium with surrounding SMA positivity and no colocalization of the two. FISH of TEI from male LGR5-EGFP enteroids implanted into female nonobese diabetic/severe combined immunodeficiency mice revealed that only the epithelium was Y chromosome positive. Chemotactic assays demonstrated increased ISEMF migration in the presence of enteroids (983 +/- 133) compared to that in the presence of either Matrigel alone (357 +/- 36) or media alone (339 +/- 24; P