title: Increased crystal-cell interaction in vitro under co-culture of renal tubular cells and adipocytes by in vitro co-culture paracrine systems simulating metabolic syndrome.
abstract: We established an experimental co-culture system for renal tubular cells and adipocytes to investigate kidney stone formation mechanisms under metabolic syndrome (MetS) conditions and examined the interaction between these cells morphologically and genetically. M-1s and 3T3-L1s were cultured individually (control, CON), with 24-h culture media from each cell type added to the other cell type (replacement, RP) in 2-layer co-culture dishes for 24 h (transwell, TW). M-1s were then exposed to calcium oxalate monohydrate (COM) crystals, and attached (14)C-labeled COM crystals were quantified. Expression of kidney stone- and adipocyte-related genes was analyzed. The radioactivity of adherent COM crystals significantly increased in TW and was relatively higher in RP compared to CON. M-1s demonstrated significant upregulation of adiponectin (Adipoq) in RP and secreted phosphoprotein 1 (Spp1) in TW compared to CON before COM crystal exposure, and significant downregulation of Spp1 in TW and upregulation of tumor necrosis factor (Tnf), interleukin 6 (Il-6), and chemokine (C-C motif) ligand 2 (Ccl2) compared to CON after COM crystal exposure. 3T3-L1s showed significant upregulation of Spp1, Adipoq, Tnf-α, and Ccl2 compared to CON. Enzyme-linked immunosorbent assays of co-culture medium revealed significantly increased TNF-α in TW. Our results highlight the potential for paracrine interactions between renal tubular cells and adipocytes and suggest that MetS conditions may lead to kidney stone formation.
authors.affiliation: Department of Nephro-urology, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya City, Aichi, 467-8601, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Ichikawa
authors.forename: Atsushi
authors.initials: A
authors.lastname: Okada
authors.forename: Kazumi
authors.initials: K
authors.lastname: Taguchi
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Fujii
authors.forename: Li
authors.initials: L
authors.lastname: Zuo
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Niimi
authors.forename: Shuzo
authors.initials: S
authors.lastname: Hamamoto
authors.forename: Yasue
authors.initials: Y
authors.lastname: Kubota
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Umemoto
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Itoh
authors.forename: Takahiro
authors.initials: T
authors.lastname: Yasui
authors.forename: Noriyasu
authors.initials: N
authors.lastname: Kawai
authors.forename: Keiichi
authors.initials: K
authors.lastname: Tozawa
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Kohri
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism*; D052242:Adiponectin / Q000235:genetics; D000818:Animals; D002129:Calcium Oxalate / Q000737:chemistry / Q000378:metabolism; D002450:Cell Communication; D002460:Cell Line; D018920:Coculture Techniques; D003460:Crystallization; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D007669:Kidney Calculi / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D007684:Kidney Tubules / Q000166:cytology* / Q000378:metabolism*; D024821:Metabolic Syndrome / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008954:Models, Biological; D053495:Osteopontin / Q000235:genetics / Q000378:metabolism; D019899:Paracrine Communication; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D052242:Adiponectin; C499919:Adipoq protein, mouse; C506896:Spp1 protein, mouse; D014409:Tumor Necrosis Factor-alpha; D053495:Osteopontin; D002129:Calcium Oxalate