The ROCK Inhibitor Y-27632 Enhances the Survival Rate of Human Embryonic Stem Cells Following Cryopreservation

2008 ◽  
Vol 17 (6) ◽  
pp. 1079-1086 ◽  
Author(s):  
Xiangyun Li ◽  
Guoliang Meng ◽  
Roman Krawetz ◽  
Shiying Liu ◽  
Derrick E. Rancourt
Keyword(s):  
Stem Cells ◽  
Survival Rate ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Rock Inhibitor

scholarly journals Novel cryopreservation method for dissociated human embryonic stem cells in the presence of a ROCK inhibitor

2008 ◽  
Vol 23 (12) ◽  
pp. 2744-2754 ◽  
Author(s):  
R. Martin-Ibanez ◽  
C. Unger ◽  
A. Stromberg ◽  
D. Baker ◽  
J.M. Canals ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Rock Inhibitor ◽  
Cryopreservation Method

scholarly journals ROCK inhibitor improves survival of cryopreserved serum/feeder-free single human embryonic stem cells

2008 ◽  
Vol 24 (3) ◽  
pp. 580-589 ◽  
Author(s):  
X. Li ◽  
R. Krawetz ◽  
S. Liu ◽  
G. Meng ◽  
D. E. Rancourt
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Rock Inhibitor

Caspase Inhibitor Z-VAD-FMK Enhances the Freeze-Thaw Survival Rate of Human Embryonic Stem Cells

2007 ◽  
Vol 27 (4-5) ◽  
pp. 257-264 ◽  
Author(s):  
Boon Chin Heng ◽  
Marie Veronique Clement ◽  
Tong Cao
Keyword(s):  
Stem Cells ◽  
Survival Rate ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Culture Media ◽  
Embryonic Stem ◽  
Immunocytochemical Staining ◽  
Irreversible Inhibitor ◽  
Slow Cooling ◽  
Spontaneous Differentiation

Previous study demonstrated that the low survival of human embryonic stem cells (hESC) under conventional slow-cooling cryopreservation protocols is predominantly due to apoptosis rather than cellular necrosis. Hence, this study investigated whether a synthetic broad-spectrum irreversible inhibitor of caspase enzymes, Z-VAD-FMK can be used to enhance the post-thaw survival rate of hESC. About 100 mM Z-VAD-FMK was supplemented into either the freezing solution, the post-thaw culture media or both. Intact and adherent hESC colonies were cryopreserved so as to enable subsequent quantitation of the post-thaw cell survival rate through the MTT assay, which can only be performed with adherent cells. Exposure to 100 mM Z-VAD-FMK in the freezing solution alone did not significantly enhance the post-thaw survival rate (10.2% vs. 9.9%, p > 0.05). However, when 100 mM Z-VAD-FMK was added to the post-thaw culture media, there was a significant enhancement in the survival rate from 9.9% to 14.4% (p < 0.05), which was further increased to 18.7% when Z-VAD-FMK was also added to the freezing solution as well (p < 0.01). Spontaneous differentiation of hESC after cryopreservation was assessed by morphological observations under bright-field microscopy, and by immunocytochemical staining for the pluripotency markers SSEA-3 and TRA-1-81. The results demonstrated that exposure to Z-VAD-FMK did not significantly enhance the spontaneous differentiation of hESC within post-thaw culture.

scholarly journals ROCK Inhibitor Is Not Required for Embryoid Body Formation from Singularized Human Embryonic Stem Cells

PLoS ONE ◽  
2014 ◽  
Vol 9 (11) ◽  
pp. e100742 ◽  
Author(s):  
Giuseppe Pettinato ◽  
Wendy S. Vanden Berg-Foels ◽  
Ning Zhang ◽  
Xuejun Wen
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Rock Inhibitor ◽  
Body Formation ◽  
Embryoid Body Formation

A ROCK inhibitor permits survival of dissociated human embryonic stem cells

2007 ◽  
Vol 25 (6) ◽  
pp. 681-686 ◽  
Author(s):  
Kiichi Watanabe ◽  
Morio Ueno ◽  
Daisuke Kamiya ◽  
Ayaka Nishiyama ◽  
Michiru Matsumura ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Rock Inhibitor

High-content screening of feeder-free human embryonic stem cells to identify pro-survival small molecules

2010 ◽  
Vol 432 (1) ◽  
pp. 21-35 ◽  
Author(s):  
Paul D. Andrews ◽  
Melissa Becroft ◽  
Anders Aspegren ◽  
Jane Gilmour ◽  
Martyn J. James ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Survival ◽  
Small Molecules ◽  
Protein Kinases ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Basic Research ◽  
Rock Inhibitor

The propensity of human embryonic stem cells to die upon enzymatic disaggregation or low-density plating is an obstacle to their isolation and routine use in drug discovery and basic research. Equally, the very low rate of establishment of implanted cells hinders cell therapy. In the present study we have developed a high-content assay for human embryonic stem cell survival and used this to screen a range of libraries of ‘lead-like’ small molecules and known bioactives. From this we identified 18 confirmed hits with four structural classes being represented by multiple compounds: a series of 5-(acyl/alkyl-amino)indazoles, compounds with a 4-(acylamino)pyridine core, simple N6,N6-dialkyladenines and compounds with a 5-(acylamino)indolinone core. In vitro kinase profiling indicated that the ROCK (Rho-associated kinase)/PRK2 (protein kinase C-related kinase 2) protein kinases are of pivotal importance for cell survival and identified previously unreported compound classes that inhibited this important biological activity. An evaluation using an extensive panel of protein kinases showed that six of our hit compounds exhibited better selectivity for ROCK inhibition than the routinely used commercially available ROCK inhibitor Y-27632. In this screen we also identified the K+-ATP channel opener pinacidil and show that it probably promotes cell survival, by ‘off-target’ inhibition of ROCK/PRK2. We have therefore identified novel pro-survival compounds of greater specificity, equivalent potency and reduced toxicity relative to the routinely employed ROCK inhibitor Y-27632.

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