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Optimization of Cryopreservation Procedures for Rat and Human Hepatocytes
Abstract
1. Rat hepatocytes were cryopreserved using a number of procedures and the viability, attachment, and metabolic activity of the cryopreserved cells were compared to freshly isolated hepatocytes. Several cryopreservation agents (dimethylsulphoxide [DMSO], glycerol, polyvinylpyrrolidone [PVP], dextrans), and combinations of these agents, were examined. Other variables tested included the freezing rate, thawing rate, and the concentration of serum in the freezing medium. 2. Recovery of viable attached cells was optimal using DMSO at concentrations of 10% or higher, a slow stepwise cooling procedure, and a quick thaw. The concentration of serum in the freezing medium (0% to 90%) did not affect cryopreservation results. Using this procedure the recovery of viable hepatocytes was 70%. 3. Levels of hepatocyte ethoxycoumarin-O-deethylase (ECOD) activity did not change following cryopreservation. The rate of decline of ECOD activity with time in culture was similar in freshly isolated and cryopreserved hepatocytes. 4. Hepatocytes isolated from three human livers were cryopreserved and recovered with viabilities similar to those obtained with the rat. A preliminary experiment also showed no loss of metabolic activity in human hepatocytes following cryopreservation.
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Cited by
Screening for human ADME/Tox drug properties in drug discovery
Li, A. P. (2001) · Drug Discovery Today · doi:10.1016/s1359-6446(01)01712-3 · Cited by 536
Self-citationLiver cell models in in vitro toxicology.
Guillouzo, A. (1998) · Environmental Health Perspectives · doi:10.1289/ehp.98106511 · Cited by 385
Li, A. P., Lu, C., Brent, J. A., Pham, C., Fackett, A., Ruegg, C. E., & Silber, P. M. (1999) · Chemico-Biological Interactions · doi:10.1016/s0009-2797(99)00088-5 · Cited by 302
Self-citationAwan, M., Buriak, I., Fleck, R., Fuller, B., Goltsev, A., Kerby, J., Lowdell, M., Mericka, P., Petrenko, A., Petrenko, Y., et al. (2020) · Regenerative Medicine · doi:10.2217/rme-2019-0145 · Cited by 253
Takahashi, H., Ayala, I., Bardet, M., De Paëpe, G., Simorre, J.-P., & Hediger, S. (2013) · Journal of the American Chemical Society · doi:10.1021/ja312501d · Cited by 149
Metadata
- Authors
- Loretz, L. J., Li, A. P., Flye, M. W., & Wilson, A. G. E.
- Venue
- Xenobiotica
- Publisher
- Informa UK Limited
- Published
- 1989-01
- Volume
- 19
- Issue
- 5
- Pages
- 489-498
- ISSN
- 0049-8254 (ISSN-L)
- 1366-5928
- OpenAlex
- W2102962539
- Cited by
- 162
- References
- 0
- FWCI
- 5.76
- Type
- journal article
- Language
- en
- Open access
- closed
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- Loretz, L. J., Li, A. P., Flye, M. W., & Wilson, A. G. E. (1989). Optimization of Cryopreservation Procedures for Rat and Human Hepatocytes. Xenobiotica. https://doi.org/10.3109/00498258909042288
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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