ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.

We report a technology to form human embryoid bodies (hEBs) from singularized human embryonic stem cells (hESCs) without the use of the p160 rho-associated coiled-coil kinase inhibitor (ROCKi) or centrifugation (spin). hEB formation was tested under four conditions: +ROCKi/+spin, +ROCKi/-spin, -ROCK...

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Main Authors: Giuseppe Pettinato, Wendy S Vanden Berg-Foels, Ning Zhang, Xuejun Wen
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC4217711?pdf=render
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spelling doaj-cdaa30b751bf4151a0da02462044be3d2020-11-25T01:46:08ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-01911e10074210.1371/journal.pone.0100742ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.Giuseppe PettinatoWendy S Vanden Berg-FoelsNing ZhangXuejun WenWe report a technology to form human embryoid bodies (hEBs) from singularized human embryonic stem cells (hESCs) without the use of the p160 rho-associated coiled-coil kinase inhibitor (ROCKi) or centrifugation (spin). hEB formation was tested under four conditions: +ROCKi/+spin, +ROCKi/-spin, -ROCKi/+spin, and -ROCKi/-spin. Cell suspensions of BG01V/hOG and H9 hESC lines were pipetted into non-adherent hydrogel substrates containing defined microwell arrays. hEBs of consistent size and spherical geometry can be formed in each of the four conditions, including the -ROCKi/-spin condition. The hEBs formed under the -ROCKi/-spin condition differentiated to develop the three embryonic germ layers and tissues derived from each of the germ layers. This simplified hEB production technique offers homogeneity in hEB size and shape to support synchronous differentiation, elimination of the ROCKi xeno-factor and rate-limiting centrifugation treatment, and low-cost scalability, which will directly support automated, large-scale production of hEBs and hESC-derived cells needed for clinical, research, or therapeutic applications.http://europepmc.org/articles/PMC4217711?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Giuseppe Pettinato
Wendy S Vanden Berg-Foels
Ning Zhang
Xuejun Wen
spellingShingle Giuseppe Pettinato
Wendy S Vanden Berg-Foels
Ning Zhang
Xuejun Wen
ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.
PLoS ONE
author_facet Giuseppe Pettinato
Wendy S Vanden Berg-Foels
Ning Zhang
Xuejun Wen
author_sort Giuseppe Pettinato
title ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.
title_short ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.
title_full ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.
title_fullStr ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.
title_full_unstemmed ROCK inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.
title_sort rock inhibitor is not required for embryoid body formation from singularized human embryonic stem cells.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2014-01-01
description We report a technology to form human embryoid bodies (hEBs) from singularized human embryonic stem cells (hESCs) without the use of the p160 rho-associated coiled-coil kinase inhibitor (ROCKi) or centrifugation (spin). hEB formation was tested under four conditions: +ROCKi/+spin, +ROCKi/-spin, -ROCKi/+spin, and -ROCKi/-spin. Cell suspensions of BG01V/hOG and H9 hESC lines were pipetted into non-adherent hydrogel substrates containing defined microwell arrays. hEBs of consistent size and spherical geometry can be formed in each of the four conditions, including the -ROCKi/-spin condition. The hEBs formed under the -ROCKi/-spin condition differentiated to develop the three embryonic germ layers and tissues derived from each of the germ layers. This simplified hEB production technique offers homogeneity in hEB size and shape to support synchronous differentiation, elimination of the ROCKi xeno-factor and rate-limiting centrifugation treatment, and low-cost scalability, which will directly support automated, large-scale production of hEBs and hESC-derived cells needed for clinical, research, or therapeutic applications.
url http://europepmc.org/articles/PMC4217711?pdf=render
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AT ningzhang rockinhibitorisnotrequiredforembryoidbodyformationfromsingularizedhumanembryonicstemcells
AT xuejunwen rockinhibitorisnotrequiredforembryoidbodyformationfromsingularizedhumanembryonicstemcells
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