title: A 3D printed microfluidic device for production of functionalized hydrogel microcapsules for culture and differentiation of human Neuronal Stem Cells (hNSC).
abstract: We present here a microfluidic device that generates sub-millimetric hollow hydrogel spheres, encapsulating cells and coated internally with a layer of reconstituted extracellular matrix (ECM) of a few microns thick. The spherical capsules, composed of alginate hydrogel, originate from the spontaneous instability of a multi-layered jet formed by co-extrusion using a coaxial flow device. We provide a simple design to manufacture this device using a DLP (digital light processing) 3D printer. Then, we demonstrate how the inner wall of the capsules can be decorated with a continuous ECM layer that is anchored to the alginate gel and mimics the basal membrane of a cellular niche. Finally, we used this approach to encapsulate human Neural Stem Cells (hNSC) derived from human Induced Pluripotent Stem Cells (hIPSC), which were further differentiated into neurons within the capsules with negligible loss of viability. Altogether, we show that these capsules may serve as cell micro-containers compatible with complex cell culture conditions and applications. These developments widen the field of research and biomedical applications of the cell encapsulation technology.
authors.affiliation: University of Geneva, Department of Biochemistry, quai Ernest Ansermet 30, CH-1211 Geneva 4, Switzerland. aurelien.roux@unige.ch and Institut Curie et Centre National de la Recherche Scientifique, Unité Mixte de Recherche 168, F-75248 Paris, France and Université Pierre et Marie Curie, F-75005 Paris, France.
authors.forename: Kevin
authors.initials: K
authors.lastname: Alessandri
authors.affiliation: Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, CH-1211 Geneva 4, Switzerland.
authors.forename: Maxime
authors.initials: M
authors.lastname: Feyeux
authors.affiliation: Institut Curie et Centre National de la Recherche Scientifique, Unité Mixte de Recherche 168, F-75248 Paris, France and Université Pierre et Marie Curie, F-75005 Paris, France and ICI, IGBMC, CNRS, UMR7104, F-67404 Illkirch-Graffenstaden, France and INSERM, U964, Université de Strasbourg, F-67400 Illkirch-Graffenstaden, France and Institut Curie et Centre National de la Recherche Scientifique, Unité Mixte de Recherche 144, F-75248 Paris, France.
authors.forename: Basile
authors.initials: B
authors.lastname: Gurchenkov
authors.affiliation: Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, CH-1211 Geneva 4, Switzerland.
authors.forename: Christophe
authors.initials: C
authors.lastname: Delgado
authors.affiliation: University of Geneva, Department of Biochemistry, quai Ernest Ansermet 30, CH-1211 Geneva 4, Switzerland. aurelien.roux@unige.ch.
authors.forename: Anastasiya
authors.initials: A
authors.lastname: Trushko
authors.affiliation: Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, CH-1211 Geneva 4, Switzerland.
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Krause
authors.affiliation: Institut Curie et Centre National de la Recherche Scientifique, Unité Mixte de Recherche 144, F-75248 Paris, France.
authors.forename: Daniela
authors.initials: D
authors.lastname: Vignjević
authors.affiliation: University of Geneva, Department of Biochemistry, quai Ernest Ansermet 30, CH-1211 Geneva 4, Switzerland. aurelien.roux@unige.ch and Institut Curie et Centre National de la Recherche Scientifique, Unité Mixte de Recherche 168, F-75248 Paris, France and Université de Bordeaux, LP2N, UMR 5298, F-33400 Talence, France and Institut d'Optique & CNRS, LP2N, UMR 5298, F-33400 Talence, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Nassoy
authors.affiliation: University of Geneva, Department of Biochemistry, quai Ernest Ansermet 30, CH-1211 Geneva 4, Switzerland. aurelien.roux@unige.ch and Swiss National Centre for Competence in Research Programme Chemical Biology, 1211 Geneva, Switzerland.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Roux
mesh_terms: D000464:Alginates / Q000737:chemistry; D057205:Automation, Laboratory; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002460:Cell Line; D002470:Cell Survival; D018914:Cells, Immobilized / Q000166:cytology* / Q000378:metabolism; D060833:Cellular Microenvironment; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D005109:Extracellular Matrix / Q000737:chemistry; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D057166:High-Throughput Screening Assays / Q000295:instrumentation; D006801:Humans; D020100:Hydrogels; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D056656:Lab-On-A-Chip Devices; D007797:Laminin / Q000737:chemistry; D009419:Nerve Tissue Proteins / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D055495:Neurogenesis; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D066330:Printing, Three-Dimensional; D000075082:Proof of Concept Study; D011509:Proteoglycans / Q000737:chemistry; D055153:Stem Cell Niche; D013499:Surface Properties
chemical_list: D000464:Alginates; D004338:Drug Combinations; D006603:Hexuronic Acids; D020100:Hydrogels; D007797:Laminin; D009419:Nerve Tissue Proteins; D011509:Proteoglycans; C059485:matrigel; D020723:Glucuronic Acid; D003094:Collagen