Gene pathway analysis of the mechanism by which the Rho-associated kinase inhibitor Y-27632 inhibits apoptosis in isolated thawed human embryonic stem cells

Ichikawa, Hinako; Nakata, N.; Abo, Youichi; Shirasawa, Sakiko; Yokoyama, Tetsuji; Yoshie, Susumu; Yue, Fengming; Tomotsune, Daihachiro; Sasaki, Katsunori

doi:10.1016/j.cryobiol.2011.11.005

Cited by 29 publications

(25 citation statements)

References 40 publications

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“…Previous studies have demonstrated that Y-27632 greatly increased the cloning efficiency of human embryonic or adult stem cells, diminished dissociation-induced apoptosis, and even enabled the cells to bypass senescence and become immortal in the presence of feeder layers. [16][17][18][19][20][21][22][23][24][25] In the present study, we found that Y-27632 significantly increased the colony-forming efficiency of limbal stem/progenitor cells, particularly the cell population with the capacity of forming colonies of large diameter. Moreover, the cloned cells maintained the expression of the putative limbal stem cell markers DNP63 and telomerase, suggesting that Y-27632 could inhibit spontaneous differentiation or even reverse the senescence of limbal stem/progenitor cells in vitro, similar to that described for human keratinocytes.…”

Section: Discussionsupporting

confidence: 51%

“…15 Y-27632, a selective inhibitor of p160-Rho-associated coiled-coil kinase, has recently been shown to increase greatly the cloning efficiency of human embryonic stem cells, keratinocytes, and corneal endothelial cells, to diminish dissociation-induced apoptosis, and even to enable cells to bypass senescence and become immortal in the presence of feeder layers. [16][17][18][19][20][21][22][23][24][25][26] In the current study, we demonstrate that the treatment of primary LECs with Y-27632 significantly increased their colony-forming efficiency by enhancing the expansion of the limbal stem/progenitor cells. Moreover, Y-27632 improved the rapid adherence of limbal stem/progenitor cells in the initial inoculation.…”

Section: Introductionmentioning

confidence: 65%

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ROCK Inhibitor Y-27632 Increases the Cloning Efficiency of Limbal Stem/Progenitor Cells by Improving Their Adherence and ROS-Scavenging Capacity

Zhou

Wang

et al. 2013

Tissue Engineering Part C: Methods

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Section: Discussionsupporting

confidence: 51%

Section: Introductionmentioning

confidence: 65%

ROCK Inhibitor Y-27632 Increases the Cloning Efficiency of Limbal Stem/Progenitor Cells by Improving Their Adherence and ROS-Scavenging Capacity

Zhou

Wang

et al. 2013

Tissue Engineering Part C: Methods

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“…RhoA GTP signaling pathways are critical for inducing several apoptotic mechanisms in response to unfavorable environmental changes (Ichikawa et al, 2013;Ohgushi et al, 2010). Inhibiting ROCK can reduce apoptosis associated with detachment and freezing protocols (Harb et al, 2008;Ichikawa et al, 2012;Koyanagi et al, 2008;Kurosawa, 2012;Olson, 2008). One of the key disadvantages of using nonviral physical delivery methods such as electroporation is the need to dissociate cells from adherent surfaces.…”

Section: Resultsmentioning

confidence: 99%

Improving Viability and Transfection Efficiency with Human Umbilical Cord Wharton's Jelly Cells Through Use of a ROCK Inhibitor

Mellott

Godsey²,

Shinogle³

et al. 2014

Cellular Reprogramming

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Differentiating stem cells using gene delivery is a key strategy in tissue engineering and regenerative medicine applications. Nonviral gene delivery bypasses several safety concerns associated with viral gene delivery; however, leading nonviral techniques, such as electroporation, subject cells to high stress and can result in poor cell viabilities. Inhibition of Rho-associated coiled-coil kinase (ROCK) has been shown to mitigate apoptotic mechanisms associated with detachment and freezing of induced pluripotent stem cells and embryonic stem cells; however, inhibiting ROCK in mesenchymal stromal cells (MSCs) for improving gene delivery applications has not been reported previously. In this study, we hypothesized that ROCK Inhibitor (RI) would improve cell viability and gene expression in primary human umbilical cord mesenchymal stromal cells (hUCMSCs) when transfected via Nucleofection™. As hypothesized, the pre-treatment and post-treatment of hUCMSCs transfected via nucleofection with Y-27632-RI significantly improved survival rates of hUCMSCs and gene expression as measured by green fluorescent protein intensity. This study provides the first comparative look at the effect of Y-27632-RI on hUCMSCs that underwent transfection via nucleofection and shows that using Y-27632-RI in concert with nucleofection could greatly enhance the utility of differentiating and reprogramming hUCMSCs for tissue engineering applications.

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“…[37][38][39][40][41][42][43][44][45][46][47][48][49][50][51] With respect to the inhibition of caspases, a variety of compounds have been shown to be effective for mammalian cells in culture. For example, P35 confers irreversible inhibition to a large number of caspases, but we chose here to focus on another popular choice in mammalian tissue culture applications, namely, Z-VAD.fmk (or its latest broad-spectrum counterpart, Q-VD-OPH) since this compound has the ability to inhibit both the intrinsic and extrinsic pathways.…”

Section: Discussionmentioning

confidence: 99%

Development of Pancreas Storage Solutions: Initial Screening of Cytoprotective Supplements for β-Cell Survival and Metabolic Status after Hypothermic Storage

Campbell

Taylor

Brockbank

2013

Biopreservation and Biobanking

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Insulin-dependent diabetes mellitus is one of the leading causes of death world-wide. Donor-derived pancreas and Islet of Langerhans transplantation are potential cures; however, postmortem ischemia impacts islet quality. The murine bt3 cell line was employed as a model to study cell viability and proliferation after hypothermic storage by comparing Belzer's Machine Perfusion Solution with UnisolÔ Solution. The objective was to determine which of these solutions provided the best base line support for bt3 cells and to screen potential cytoprotective additives to the solutions. Initial bt3 cell viability was similar in the two storage solutions; however, better proliferation was observed after storage in Unisol Solution. The caspase inhibitor, Q-VD-OPH, and a-tocopherol improved viability in both storage solutions, suggesting that apoptotic pathways may be responsible for cell death during hypothermic storage of bt3 cells. Analysis of apoptosis markers, caspase activity, and DNA laddering showed a reduction in apoptosis when these additives were included. The effects of Q-VD-OPH and a-tocopherol were also synergistic when employed together during either hypothermic exposure, post-hypothermic physiologic incubation, or combinations of hypothermic exposure and physiologic incubation. These results suggest that both supplements should be included in pancreas hypothermic storage solutions and in islet culture media during post-isolation culture prior to transplantation.

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Gene pathway analysis of the mechanism by which the Rho-associated kinase inhibitor Y-27632 inhibits apoptosis in isolated thawed human embryonic stem cells

Cited by 29 publications

References 40 publications

ROCK Inhibitor Y-27632 Increases the Cloning Efficiency of Limbal Stem/Progenitor Cells by Improving Their Adherence and ROS-Scavenging Capacity

ROCK Inhibitor Y-27632 Increases the Cloning Efficiency of Limbal Stem/Progenitor Cells by Improving Their Adherence and ROS-Scavenging Capacity

Improving Viability and Transfection Efficiency with Human Umbilical Cord Wharton's Jelly Cells Through Use of a ROCK Inhibitor

Development of Pancreas Storage Solutions: Initial Screening of Cytoprotective Supplements for β-Cell Survival and Metabolic Status after Hypothermic Storage

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