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

Sergey Rodin, Liselotte Antonsson, Colin Niaudet, Oscar E. Simonson, Elina Salmela, Emil M. Hansson, Anna Domogatskaya, Zhijie Xiao, Pauliina Damdimopoulou, Mona Sheikhi, José Inzunza, Ann-Sofie Nilsson, Duncan Baker, Raoul Kuiper, Yi Sun, Elisabeth Blennow, Magnus Nordenskjöld, Karl-Henrik Grinnemo, Juha Kere, Christer Betsholtz, Outi Hovatta and Karl Tryggvason ()
Additional contact information
Sergey Rodin: Karolinska Institute
Liselotte Antonsson: Intervention and Technology, Karolinska Institute and Karolinska University Hospital, Huddinge
Colin Niaudet: Vascular Biology, Karolinska Institute
Oscar E. Simonson: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institute and Karolinska University Hospital
Elina Salmela: Research Programs Unit, Molecular Neurology, University of Helsinki
Emil M. Hansson: Cardiovascular Research Center, Massachusetts General Hospital, Charles River Plaza/CPZN 3200, 185 Cambridge Street
Anna Domogatskaya: Karolinska Institute
Zhijie Xiao: Karolinska Institute
Pauliina Damdimopoulou: Intervention and Technology, Karolinska Institute and Karolinska University Hospital, Huddinge
Mona Sheikhi: Intervention and Technology, Karolinska Institute and Karolinska University Hospital, Huddinge
José Inzunza: Intervention and Technology, Karolinska Institute and Karolinska University Hospital, Huddinge
Ann-Sofie Nilsson: Karolinska Institute
Duncan Baker: Sheffield Diagnostic Genetic Services, Sheffield Children’s NHS Trust
Raoul Kuiper: FENO, Karolinska Institute, Huddinge, Karolinska Hospital
Yi Sun: BioLamina AB, Löfströms Allé 5A
Elisabeth Blennow: Karolinska University Hospital
Magnus Nordenskjöld: Karolinska University Hospital
Karl-Henrik Grinnemo: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institute and Karolinska University Hospital
Juha Kere: Research Programs Unit, Molecular Neurology, University of Helsinki
Christer Betsholtz: Vascular Biology, Karolinska Institute
Outi Hovatta: Intervention and Technology, Karolinska Institute and Karolinska University Hospital, Huddinge
Karl Tryggvason: Karolinska Institute

Nature Communications, 2014, vol. 5, issue 1, 1-13

Abstract: 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.

Date: 2014
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DOI: 10.1038/ncomms4195

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