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Review
. 2010 Sep;5(5):799-807.
doi: 10.2217/rme.10.45.

Stimulating progress in regenerative medicine: improving the cloning and recovery of cryopreserved human pluripotent stem cells with ROCK inhibitors

Angie Rizzino  1
Affiliations
Review

Stimulating progress in regenerative medicine: improving the cloning and recovery of cryopreserved human pluripotent stem cells with ROCK inhibitors

Angie Rizzino. Regen Med. 2010 Sep.

Abstract

Until recently, culturing human pluripotent stem cells was hampered by three prominent technical problems: a high degree of unwanted cellular stress when the cells are passaged, unacceptably low cloning efficiency and poor recovery of cryopreserved stocks. This review discusses recent developments that address these problems. A major focus of the review is the use of p160 Rho-associated coiled-coil kinase inhibitors for improving both the cloning efficiency and the recovery of cryopreserved human embryonic stem cells and human induced pluripotent stem cells. An underlying theme of this review is that the three problems have a common cause: separation of human pluripotent stem cells from one another increases cellular stress, which greatly decreases their viability unless special steps are taken.

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Figures

Figure 1
Figure 1. Rho-associated coiled-coil kinase signaling pathway
The p160 ROCK is activated by several upstream pathways, including activated Rho-GTP and activated caspase 3. ROCK has a wide range of targets that are regulated when phosphorylated by ROCK. Downstream signaling of ROCK can influence many different biological responses, including cell migration, cell adhesion and apoptosis. ROCKi are believed to primarily inhibit the activity of ROCK 1 and 2, but may influence the activity of other kinases. ERM: Ezrin, radixin and moesin; GF: Growth factor; GFAP: Glial fibrillary acidic protein; GFR: Growth factor receptor; GPCR: G-protein-coupled receptor; MLC: Myosin light chain; MLCP: Myosin light chain phosphatase; NF-L: Neurofilament protein; NHE: Na+/H+ exchanger; PRK: Protein kinase C-related kinase; ROCK: Rho-associated coiled-coil kinase; ROCKi: ROCK inhibitor.
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References

    1. Thomson JA, Itskovitz-Eldor J, Shapiro SS, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998;282(5395):1145–1147. First report demonstrating the isolation of human embryonic stem cells (hESCs) - PubMed
    1. Takahashi K, Tanabe K, Ohnuki M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131(5):861–872. - PubMed
    1. Wernig M, Meissner A, Foreman R, et al. In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature. 2007;448(7151):318–324. - PubMed
    1. Yu J, Vodyanik MA, Smuga-Otto K, et al. Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007;318(5858):1917–1920. - PubMed
    1. Kiskinis E, Eggan K. Progress toward the clinical application of patient-specific pluripotent stem cells. J Clin Invest. 2010;120(1):51–59. - PMC - PubMed

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