R Bücker1, C Schaefer2, A D Gruber3, J Hoppe3, L Lazzerini2, J Barinoff2, J Sehouli2, Günter Cichon4. 1. Department of Gastroenterology, Institute of Clinical Physiology, Charité - Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany. 2. Department of Gynecology, Charite-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany. 3. Institute of Veterinary Pathology, Free University Berlin, Berlin, Germany. 4. Department of Gynecology, Charite-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany. guenter.cichon@charite.de.
Abstract
PURPOSE: The identification of novel cell lines which combine the most important properties of mucosal membranes in terms of drug absorption, transmembrane transport and mucus secretion can help to establish improved and meaningful test systems for pharmacological and infectiological studies. METHODS: We have established a novel mucus secreting tumor cell line (Cx-03) derived from a female patient who underwent radical hysterectomy after diagnosis of a large malignant carcino sarcoma (Muellerian mixed tumor). Via xenotransplantation in SCID beige mice, recultivation and subcloning a stable cell line was established from primary tumor cells. RESULTS: Human origin and novelty of the cell line was determined by karyotype analysis and STR fingerprint. During growth cells produce considerable amounts of a PAS positive viscoelastic mucus. Immunostaining revealed expression of mucins and the mucin modifier CLCA1. We demonstrate in initial electrophysiological experiments that confluent, polarized monolayers of Cx-03 are formed (on PCF-filter supports) that exhibit stable electrical resistance (> 600 Ω cm2). Confluent Cx-03 monolayers express barrier-forming tight junction proteins claudin-1 and -4 which co-localize with zonula occludens protein-1 (ZO-1) at cell-cell contacts. CONCLUSIONS: Mucus secretion is a rare property among mammalian cell lines. In combination with its ability to form polarized monolayers Cx-03 might contribute as a novel cell based model for drug absorption, transport and barrier studies.
PURPOSE: The identification of novel cell lines which combine the most important properties of mucosal membranes in terms of drug absorption, transmembrane transport and mucus secretion can help to establish improved and meaningful test systems for pharmacological and infectiological studies. METHODS: We have established a novel mucus secreting tumor cell line (Cx-03) derived from a female patient who underwent radical hysterectomy after diagnosis of a large malignant carcino sarcoma (Muellerian mixed tumor). Via xenotransplantation in SCID beige mice, recultivation and subcloning a stable cell line was established from primary tumor cells. RESULTS:Human origin and novelty of the cell line was determined by karyotype analysis and STR fingerprint. During growth cells produce considerable amounts of a PAS positive viscoelastic mucus. Immunostaining revealed expression of mucins and the mucin modifier CLCA1. We demonstrate in initial electrophysiological experiments that confluent, polarized monolayers of Cx-03 are formed (on PCF-filter supports) that exhibit stable electrical resistance (> 600 Ω cm2). Confluent Cx-03 monolayers express barrier-forming tight junction proteins claudin-1 and -4 which co-localize with zonula occludens protein-1 (ZO-1) at cell-cell contacts. CONCLUSIONS: Mucus secretion is a rare property among mammalian cell lines. In combination with its ability to form polarized monolayers Cx-03 might contribute as a novel cell based model for drug absorption, transport and barrier studies.
Entities:
Keywords:
CLCA1; MUC2; barrier function; mucus screting cell line
Authors: E Gullberg; M Leonard; J Karlsson; A M Hopkins; D Brayden; A W Baird; P Artursson Journal: Biochem Biophys Res Commun Date: 2000-12-29 Impact factor: 3.575
Authors: José das Neves; Cristina M R Rocha; Maria Pilar Gonçalves; Rebecca L Carrier; Mansoor Amiji; Maria Fernanda Bahia; Bruno Sarmento Journal: Mol Pharm Date: 2012-10-08 Impact factor: 4.939
Authors: José das Neves; Francisca Araújo; Fernanda Andrade; Johan Michiels; Kevin K Ariën; Guido Vanham; Mansoor Amiji; Maria Fernanda Bahia; Bruno Sarmento Journal: Mol Pharm Date: 2013-06-17 Impact factor: 4.939