title: [Culture supernatants of lymphocytes from different lymphoid tissues induce transdifferentiation of Caco2 cells into M-like cells].
abstract: OBJECTIVE
  To establish the microfold (M)-like cell model in vitro and identify M-like cells through detecting the capacity of transporting fluorescent beads and the levels of the associated genes, and to observe the effects of lymphocyte culture supernatants stimulated by concanavalin A (Con A) from different lymphoid tissues on the differentiation of Caco2 cells into M-like cells.
  
  
  METHODS
  The isolated lymphocytes of Peyer's patch (PP), mesenteric lymph node (MLN) and spleen (Sp) were incubated with 3 μg/mL Con A for 3 days. The culture supernatants were collected and co-cultured with Caco2 cells. The fluorescent bead suspension was added into the upper compartment of the Transwell™ inserts, and then basolateral solutions were then sampled and analyzed. The number of transported fluorescent beads was measured by flow cytometry. The expressions of M-like cells-associated genes, such as chemokine (C-C motif) ligand 20 (CCL20), claudin4 (CLDN4), tumor necrosis factor receptor superfamily member 9 (TNFRSF9), and Spi-B were detected by reverse transcription PCR.
  
  
  RESULTS
  Compared with blank control group, the number of fluorescent beads transported by induced Caco2 cells and the levels of CCL20, CLDN4, TNFRSF9 and Spi-B mRNAs significantly increased in induced Caco2 cells treated with the culture supernatants of lymphocytes from PP, MLN and Sp. After Con A stimulation, the number of fluorescent beads transported by induced Caco2 cells and the levels of CCL20, CLDN4, TNFRSF9 and Spi-B mRNAs were higher than those in the unstimulated group.
  
  
  CONCLUSIONS
  The lymphocyte culture supernatants stimulated or unstimulated by Con A can induce the transdifferentiation of Caco2 cells into M-like cells.
authors.affiliation: Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Wang
authors.affiliation: Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Luo
authors.affiliation: Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
authors.forename: Dongsheng
authors.initials: D
authors.lastname: Zheng
authors.affiliation: Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
authors.forename: Lian
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
authors.forename: Yuanhong
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Guangzhou University of Chinese Medicine, Guangzhou 510405, China.
authors.forename: Xiu
authors.initials: X
authors.lastname: Wu
mesh_terms: D000818:Animals; D018938:Caco-2 Cells; D054338:Cell Transdifferentiation; D054418:Chemokine CCL20 / Q000235:genetics; D062506:Claudin-4 / Q000235:genetics; D003208:Concanavalin A / Q000494:pharmacology; D006801:Humans; D008214:Lymphocytes / Q000502:physiology*; D008221:Lymphoid Tissue / Q000166:cytology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010581:Peyer's Patches / Q000166:cytology*; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000235:genetics
chemical_list: C108355:CCL20 protein, human; C568157:CLDN4 protein, human; D054418:Chemokine CCL20; D062506:Claudin-4; C505714:TNFRSF9 protein, human; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; D003208:Concanavalin A