title: Facile bench-top fabrication of enclosed circular microchannels provides 3D confined structure for growth of prostate epithelial cells.
abstract: We present a simple bench-top method to fabricate enclosed circular channels for biological experiments. Fabricating the channels takes less than 2 hours by using glass capillaries of various diameters (from 100 µm up to 400 µm) as a mould in PDMS. The inner surface of microchannels prepared in this way was coated with a thin membrane of either Matrigel or a layer-by-layer polyelectrolyte to control cellular adhesion. The microchannels were then used as scaffolds for 3D-confined epithelial cell culture. To show that our device can be used with several epithelial cell types from exocrine glandular tissues, we performed our biological studies on adherent epithelial prostate cells (non-malignant RWPE-1 and invasive PC3) and also on breast (non-malignant MCF10A) cells We observed that in static conditions cells adhere and proliferate to form a confluent layer in channels of 150 µm in diameter and larger, whereas cellular viability decreases with decreasing diameter of the channel. Matrigel and PSS (poly (sodium 4-styrenesulphonate)) promote cell adhesion, whereas the cell proliferation rate was reduced on the PAH (poly (allylamine hydrochloride))-terminated surface. Moreover infusing channels with a continuous flow did not induce any cellular detachment. Our system is designed to simply grow cells in a microchannel structure and could be easily fabricated in any biological laboratory. It offers opportunities to grow epithelial cells that support the formation of a light. This system could be eventually used, for example, to collect cellular secretions, or study cell responses to graduated hypoxia conditions, to chemicals (drugs, siRNA, …) and/or physiological shear stress.
authors.affiliation: Univ. Grenoble Alpes, iRTSV-BGE, Grenoble, France; CEA, iRTSV-BGE, Grenoble, France; INSERM, BGE, Grenoble, France.
authors.forename: Monika E
authors.initials: ME
authors.lastname: Dolega
authors.affiliation: Univ. Grenoble Alpes, iRTSV-BGE, Grenoble, France; CEA, iRTSV-BGE, Grenoble, France; INSERM, BGE, Grenoble, France.
authors.forename: Jayesh
authors.initials: J
authors.lastname: Wagh
authors.affiliation: Univ. Grenoble Alpes, iRTSV-BGE, Grenoble, France; CEA, iRTSV-BGE, Grenoble, France; INSERM, BGE, Grenoble, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Gerbaud
authors.affiliation: Univ. Grenoble Alpes, iRTSV-BGE, Grenoble, France; CEA, iRTSV-BGE, Grenoble, France; INSERM, BGE, Grenoble, France.
authors.forename: Frederique
authors.initials: F
authors.lastname: Kermarrec
authors.affiliation: UJF-Grenoble 1, CNRS, TIMC-IMAG UMR 5525 (SyNaBi), Grenoble, France.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Alcaraz
authors.affiliation: UJF-Grenoble 1, CNRS, TIMC-IMAG UMR 5525 (SyNaBi), Grenoble, France.
authors.forename: Donald K
authors.initials: DK
authors.lastname: Martin
authors.affiliation: Univ. Grenoble Alpes, iRTSV-BGE, Grenoble, France; CEA, iRTSV-BGE, Grenoble, France; INSERM, BGE, Grenoble, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: Gidrol
authors.affiliation: Univ. Grenoble Alpes, iRTSV-BGE, Grenoble, France; CEA, iRTSV-BGE, Grenoble, France; INSERM, BGE, Grenoble, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Picollet-D'hahan
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects*; D018929:Cell Culture Techniques; D015687:Cell Hypoxia / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000008:administration & dosage; D004338:Drug Combinations; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D007797:Laminin / Q000008:administration & dosage; D008297:Male; D011073:Polyamines / Q000008:administration & dosage; D011467:Prostate / Q000166:cytology* / Q000187:drug effects; D011509:Proteoglycans / Q000008:administration & dosage; D023822:Tissue Engineering
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011073:Polyamines; D011509:Proteoglycans; C059485:matrigel; C063994:polyallylamine; D003094:Collagen