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

• 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.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinsk bioteknologi -- Biomedicinsk laboratorievetenskap/teknologi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Medical Biotechnology -- Biomedical Laboratory Science/Technology (hsv//eng)

Nyckelord

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

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