title: Efficient passage of human pluripotent stem cells on spider silk matrices under xeno-free conditions.
abstract: Human pluripotent stem cells (hPSCs) hold great promise for applications in regenerative medicine and pharmaceutical development. Such applications require cell culture methods and reagents that are chemically defined, xeno-free, scalable, and low-cost. Herein, we describe non-mechanical passaging of hPSCs on spider silk films under chemically defined and xeno-free conditions. The cells were dissociated into single cells or small aggregates using Accutase or enzyme-free dissociation buffer and then passaged to spider silk films, where they expanded in monolayers until they covered the surface. Cells cultured over 10 passages on spider silk film remained karyotypically normal and pluripotent. In conclusion, a novel method for passaging dissociated hPSCs under conditions that are compatible with clinical applications is presented. The method is cost-efficient and may be useful for both research and clinical applications.
authors.affiliation: Division of Neurogeriatrics, Department of Neurobiology, Care Sciences and Society (NVS), Center for Alzheimer Research, Karolinska Institutet, Huddinge, 14157, Stockholm, Sweden.
authors.forename: Siqin
authors.initials: S
authors.lastname: Wu
authors.affiliation: Division of Neurogeriatrics, Department of Neurobiology, Care Sciences and Society (NVS), Center for Alzheimer Research, Karolinska Institutet, Huddinge, 14157, Stockholm, Sweden.|Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, Box 7011, 75007, Uppsala, Sweden.|Institute of Mathematics and Natural Sciences, Tallinn University, Narva mnt 25, 10120, Tallinn, Estonia.
authors.forename: Jan
authors.initials: J
authors.lastname: Johansson
authors.affiliation: Division of Obstetrics and Gynecology, Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet and Karolinska University Hospital, Huddinge, 14186, Stockholm, Sweden.
authors.forename: Outi
authors.initials: O
authors.lastname: Hovatta
authors.affiliation: Division of Neurogeriatrics, Department of Neurobiology, Care Sciences and Society (NVS), Center for Alzheimer Research, Karolinska Institutet, Huddinge, 14157, Stockholm, Sweden. Anna.Rising@ki.se.|Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, Box 7011, 75007, Uppsala, Sweden. Anna.Rising@ki.se.
authors.forename: Anna
authors.initials: A
authors.lastname: Rising
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005136:Eye Proteins / Q000378:metabolism; D051542:Hepatocyte Nuclear Factor 3-beta / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D007621:Karyotyping; D008856:Microscopy, Fluorescence; D000071317:Nanog Homeobox Protein; D064231:Nestin / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D012097:Repressor Proteins / Q000378:metabolism; D055760:SOXF Transcription Factors / Q000378:metabolism; D047011:Silk / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D013112:Spiders / Q000378:metabolism
chemical_list: D005136:Eye Proteins; C496502:FOXA2 protein, human; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D064231:Nestin; D050814:Octamer Transcription Factor-3; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; C494173:POU5F1 protein, human; D051761:Paired Box Transcription Factors; D011994:Recombinant Proteins; D012097:Repressor Proteins; C109188:SOX17 protein, human; D055760:SOXF Transcription Factors; D047011:Silk; D051542:Hepatocyte Nuclear Factor 3-beta
keywords: Culture matrices; Differentiation; Enzymatic passaging; Self-renewal; hESC; iPSC