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Nat Commun. 2014;5:3195. doi: 10.1038/ncomms4195.

Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment.

Author information

1
Division of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm SE-171 77, Sweden.
2
Division of Obstetrics and Gynecology, Department of Clinical Sciences, Intervention and Technology, Karolinska Institute and Karolinska University Hospital, Huddinge, Stockholm SE-141 86, Sweden.
3
Vascular Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute Stockholm SE-171 77, Sweden.
4
Division of Cardiothoracic Surgery and Anaesthesiology, Department of Molecular Medicine and Surgery, Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institute and Karolinska University Hospital, Stockholm SE-17177, Sweden.
5
1] Research Programs Unit, Molecular Neurology, University of Helsinki, Helsinki FI-00014, Finland [2] Folkhälsan Institute of Genetics, Helsinki FI-00014, Finland.
6
Cardiovascular Research Center, Massachusetts General Hospital, Charles River Plaza/CPZN 3200, 185 Cambridge Street, Boston, Massachusetts 02114-2790, USA.
7
Sheffield Diagnostic Genetic Services, Sheffield Children's NHS Trust, Sheffield S102TH, UK.
8
Department of Laboratory Medicine, FENO, Karolinska Institute, Huddinge, Karolinska Hospital, Huddinge, Stockholm SE-141 86, Sweden.
9
BioLamina AB, Löfströms Allé 5A, Sundbyberg 17266, Sweden.
10
Department of Molecular Medicine and Surgery and Center for Molecular Medicine Karolinska Institutet, Karolinska University Hospital, Stockholm 171 76, Sweden.
11
1] Research Programs Unit, Molecular Neurology, University of Helsinki, Helsinki FI-00014, Finland [2] Folkhälsan Institute of Genetics, Helsinki FI-00014, Finland [3] Department of Biosciences and Nutrition and Science for Life Laboratory, Karolinska Institute, Stockholm SE-141 83 Sweden.
12
1] Division of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm SE-171 77, Sweden [2] Cardiovascular and Metabolic Disorders Program, Duke-NUS, Singapore City 169857, Singapore.

Abstract

Lack of robust methods for establishment and expansion of pluripotent human embryonic stem (hES) cells still hampers development of cell therapy. Laminins (LN) are a family of highly cell-type specific basement membrane proteins important for cell adhesion, differentiation, migration and phenotype stability. Here we produce and isolate a human recombinant LN-521 isoform and develop a cell culture matrix containing LN-521 and E-cadherin, which both localize to stem cell niches in vivo. This matrix allows clonal derivation, clonal survival and long-term self-renewal of hES cells under completely chemically defined and xeno-free conditions without ROCK inhibitors. Neither LN-521 nor E-cadherin alone enable clonal survival of hES cells. The LN-521/E-cadherin matrix allows hES cell line derivation from blastocyst inner cell mass and single blastomere cells without a need to destroy the embryo. This method can facilitate the generation of hES cell lines for development of different cell types for regenerative medicine purposes.

PMID:
24463987
DOI:
10.1038/ncomms4195
[Indexed for MEDLINE]
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