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Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality

Human embryonic stemcells (hESCs) and induced pluripotent stemcells (iPSCs) provide an unlimited source for the generation of human hepatocytes, owing to their indefinite self-renewal and pluripotent properties. Both hESC-/iPSC-derived hepatocytes hold great promise in treating liver diseases as pot...

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Main Authors: Ramasamy, T.S., Yu, J.S.L., Selden, C., Hodgson, H., Cui, W.
Format: Article
Language:English
Published: 2013
Subjects:
R Medicine
Online Access:http://eprints.um.edu.my/9241/1/TS_ramasamy_tissue_engineering_2013.pdf
http://eprints.um.edu.my/9241/
http://online.liebertpub.com/doi/abs/10.1089/ten.tea.2012.0190
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spelling my.um.eprints.92412014-02-07T02:49:34Z http://eprints.um.edu.my/9241/ Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality Ramasamy, T.S. Yu, J.S.L. Selden, C. Hodgson, H. Cui, W. R Medicine Human embryonic stemcells (hESCs) and induced pluripotent stemcells (iPSCs) provide an unlimited source for the generation of human hepatocytes, owing to their indefinite self-renewal and pluripotent properties. Both hESC-/iPSC-derived hepatocytes hold great promise in treating liver diseases as potential candidates for cell replacement therapies or as an in vitro platform to conduct newdrug trials. It has been previously demonstrated that the initiation of hESC differentiation in monolayer cultures increases the generation of definitive endoderm (DE) and subsequently of hepatocyte differentiation. However, monolayer culture may hinder the maturation of hESC-derived hepatocytes, since such two-dimensional (2D) conditions do not accurately reflect the complex nature of three-dimensional (3D) hepatocyte specification in vivo. Here, we report the sequential application of 2D and 3D culture systems to differentiate hESCs to hepatocytes. Human ESCs were initially differentiated in a monolayer culture to DE cells, which were then inoculated into Algimatrix scaffolds. Treatments of hESC-DE cells with a ROCK inhibitor before and after inoculation dramatically enhanced their survival and the formation of spheroids, which are distinct from HepG2 carcinoma cells. In comparison with monolayer culture alone, sequential 2D and 3D cultures significantly improved hepatocyte differentiation and function. Our results demonstrate that hESC-DE cells can be incorporated into Algimatrix 3D culture systems to enhance hepatocyte differentiation and function. 2013 Article PeerReviewed application/pdf en http://eprints.um.edu.my/9241/1/TS_ramasamy_tissue_engineering_2013.pdf Ramasamy, T.S. and Yu, J.S.L. and Selden, C. and Hodgson, H. and Cui, W. (2013) Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality. Tissue Engineering Part A, 19 (3-4). pp. 360-367. ISSN 1937-3341 http://online.liebertpub.com/doi/abs/10.1089/ten.tea.2012.0190 10.1089/ten.tea.2012.0190
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
language English
topic R Medicine
spellingShingle R Medicine
Ramasamy, T.S.
Yu, J.S.L.
Selden, C.
Hodgson, H.
Cui, W.
Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality
description Human embryonic stemcells (hESCs) and induced pluripotent stemcells (iPSCs) provide an unlimited source for the generation of human hepatocytes, owing to their indefinite self-renewal and pluripotent properties. Both hESC-/iPSC-derived hepatocytes hold great promise in treating liver diseases as potential candidates for cell replacement therapies or as an in vitro platform to conduct newdrug trials. It has been previously demonstrated that the initiation of hESC differentiation in monolayer cultures increases the generation of definitive endoderm (DE) and subsequently of hepatocyte differentiation. However, monolayer culture may hinder the maturation of hESC-derived hepatocytes, since such two-dimensional (2D) conditions do not accurately reflect the complex nature of three-dimensional (3D) hepatocyte specification in vivo. Here, we report the sequential application of 2D and 3D culture systems to differentiate hESCs to hepatocytes. Human ESCs were initially differentiated in a monolayer culture to DE cells, which were then inoculated into Algimatrix scaffolds. Treatments of hESC-DE cells with a ROCK inhibitor before and after inoculation dramatically enhanced their survival and the formation of spheroids, which are distinct from HepG2 carcinoma cells. In comparison with monolayer culture alone, sequential 2D and 3D cultures significantly improved hepatocyte differentiation and function. Our results demonstrate that hESC-DE cells can be incorporated into Algimatrix 3D culture systems to enhance hepatocyte differentiation and function.
format Article
author Ramasamy, T.S.
Yu, J.S.L.
Selden, C.
Hodgson, H.
Cui, W.
author_facet Ramasamy, T.S.
Yu, J.S.L.
Selden, C.
Hodgson, H.
Cui, W.
author_sort Ramasamy, T.S.
title Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality
title_short Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality
title_full Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality
title_fullStr Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality
title_full_unstemmed Application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality
title_sort application of three-dimensional culture conditions to human embryonic stem cell-derived definitive endoderm cells enhances hepatocyte differentiation and functionality
publishDate 2013
url http://eprints.um.edu.my/9241/1/TS_ramasamy_tissue_engineering_2013.pdf
http://eprints.um.edu.my/9241/
http://online.liebertpub.com/doi/abs/10.1089/ten.tea.2012.0190
_version_ 1643688509615112192
score 13.211869

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