Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/81956
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Type: Journal article
Title: Decellularized feeders: an optimized method for culturing pluripotent cells
Author: Lim, M.
Jungebluth, P.
Sjoqvist, S.
Nidkin, H.
Kjartansdottir, K.
Unger, C.
Vassiliev, I.
Macchiarini, P.
Citation: Stem Cells Translational Medicine, 2013; 2(12):975-982
Publisher: AlphaMed Press, Inc
Issue Date: 2013
ISSN: 2157-6564
2157-6580
Statement of
Responsibility: 
Mei Ling Lim, Philipp Jungebluth, Sebastian Sjöqvist, Hero Nikdin, Kristín Rós Kjartansdóttir, Christian Unger, Ivan Vasslieve and Paolo Macchiarini
Abstract: Pluripotent cells such as human embryonic stem cells and human induced pluripotent stem cells are useful in the field of regenerative medicine because they can proliferate indefinitely and differentiate into all cell types. However, a limiting factor for maintaining and propagating stem cells is the need for inactivated fibroblasts as a growth matrix, since these may potentially cause cross-contamination. In this study, we aimed to maintain stem cells on the extracellular matrix (ECM) of either nonirradiated or γ-irradiated fibroblasts. It has been demonstrated that the ECM contains factors and proteins vital for the adhesion, proliferation, and differentiation of pluripotent cells. In order to preserve the ECM, the cell layers of the fibroblasts were decellularized by treatment with 0.05% sodium dodecyl sulfate (SDS), which resulted in an absence of DNA as compared with conventional feeder culture. However, SDS treatment did not cause a detectable change in the ECM architecture and integrity. Furthermore, immunohistochemistry demonstrated that expressions of major ECM proteins, such as fibronectin, collagen, and laminin, remained unaltered. The human pluripotent cells cultured on this decellularized matrix maintained gene expression of the pluripotency markers NANOG and OCT4 and had the potency to differentiate to three germ layers. The in vitro culture system shown here has an excellent potential since the main allogeneic components (i.e., DNA of the feeder cells) are removed. It is also a technically easy, fast, safe, and cheap method for maintaining a refined feeder-free stem cell culture for further cell differentiation studies.
Keywords: Pluripotent stem cells
Technology
Scaffold attachment region
Cell culture
Serum-free media
Stem cell
Stem cell culture
Extracellular matrix
Rights: ©AlphaMed Press
DOI: 10.5966/sctm.2013-0077
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