Cell line/type | ESCs (Embryonic Stem Cells) |
---|---|
Species | Human |
Animal free | No |
Product | KSR-DMSO + Y-27632 (cryopreservation) |
Li, X., Krawetz, R., Liu, S., Meng, G., & Rancourt, D. E. (2008). ROCK inhibitor improves survival of cryopreserved serum/feeder-free single human embryonic stem cells. Human reproduction, 24(3), 580-589. In his study, hES cells maintained on Matrigel-coated dishes were dissociated into single cells with Accutase and slow freezing. After thawing at 37 degrees C, cells were cultured in mTeSR medium supplemented with 10 microM of Rho-associated kinase inhibitor Y-27632 for 1 day. The use of Y-27632 and Accutase significantly increases the survival of single hES cells after thawing compared with a control group. Furthermore, by treatment of hES cell aggregates with EGTA to disrupt cell-cell interaction, the authors showed that Y-27632 treatment does not directly affect hES cell apoptosis. Even in the presence of Y-27632, hES cells deficient in cell-cell interaction undergo apoptosis. Y-27632-treated freeze-thawed hES cells retain typical morphology, stable karyotype, expression of pluripotency markers and the potential to differentiate into derivatives of all three germ layers after long-term culture. The method described here allows for cryopreservation of single hES cells in serum-free and feeder-free conditions and therefore we believe this method will be ideal for current and future hES cell applications that are targeted towards a therapeutic end-point. The freezing medium consisting of 90% knockout serum replacement and 10% DMSO, supplemented with Y-27632 as specified in the Materials and Methods, under the heading; Freezing and thawing of hES cells. https://www.ncbi.nlm.nih.gov/pubmed/19056770 |
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Source | Literature - modified commercial product |
Chemically defined > Yes | Yes |
Contains phenol red > No | No |
Antibiotics free > Yes | Yes |