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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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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