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Upregulation of adhesion molecules sustains matrix-free growth of human embryonic stem cells

Journal article
Authors Narmin Bigdeli
Giuseppe Maria de Peppo
Camilla Karlsson
Maria Lennerås
R. Strehl
J. Hyllner
Anders Lindahl
Published in Open Stem Cell Journal
Volume 5
Issue 1
Pages 14-30
ISSN 1876-8938
Publication year 2018
Published at Institute of Biomedicine, Department of Clinical Chemistry and Transfusion Medicine
Institute of Clinical Sciences, Department of Biomaterials
Pages 14-30
Language en
Keywords Attachment proteins, Cell therapy, Human embryonic stem cells, Integrins, Matrix-free culture, Microarray analysis, Regenerative medicine, RGD-associated proteins, arginylglycylaspartic acid, beta5 integrin, caveolin 1, cell adhesion molecule, deah box helicase 9, deoxyribonuclease, fibronectin, heterogeneous nuclear ribonucleoprotein group F H, heterogeneous nuclear ribonucleoprotein U, initiation factor 2, initiation factor 3, max interacting protein 2, nonstructural protein 1, nucleoporin, nucleoporin 100, nucleoporin 133, ribosome protein, ribosome protein 27a, ribosome protein l37a, ribosome protein lateral stalk subunit 38, ribosome protein lateral stalk subunit p2, ribosome protein s11, ribosome protein s15, RNA binding protein FUS, RNA splicing factor, splicing factor 3b subunit 1, transcription factor NANOG, unclassified drug, unindexed drug, Article, cell adhesion, cell differentiation, cell growth, controlled study, extracellular matrix, flow cytometry, gene expression, gene overexpression, human, human cell, oncogene, phase contrast microscopy, priority journal, protein interaction, real time polymerase chain reaction, reverse transcription polymerase chain reaction, RNA isolation, upregulation
Subject categories Biomedical Laboratory Science/Technology


Background: Despite recent advances in culture techniques for undifferentiated human Embryonic Stem Cells (hESCs), further improvements are required to facilitate research and translation of these cells in clinical settings. We have previously derived hESC lines that can be cultured in their undifferentiated state on regular plastic culture dishes, without the need for feeder cells or other coating supports, denoted Matrix-Free Growth hESCs (MFG-hESCs). Objective: In this study, we further characterize and compare MFG-hESCs to hESCs in order to understand the molecular differences responsible for the unique ability of MFG-hESCs. Results: Microarray analysis demonstrated that MFG-hESCs highly resemble feeder-cultured hESCs in global gene expression profile. Two identified groups of genes with differential expression were those encoding for ribosomal proteins and attachment proteins, such as the RGD (Arg-Gly-Asp)-associated proteins. Real-time PCR and flow cytometry corroborated the microarray results. Culture of MFG-hESCs in the presence of RGD peptides resulted in decreased attachment ability compared to cells cultured in the presence of RGES (Arg-Gly-Asp-Ser) peptides. Conclusion: This study demonstrates that MFG-hESC lines overexpress cell attachment proteins but retain the typical characteristics of undifferentiated feeder-cultured hESCs. The ability to culture high-quality pluripotent stem cells in feeder-and matrix-free conditions creates a new opportunities for their large-scale manufacturing for experimental research and translational applications. © 2018 Bigdeli et al.

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