Cryoprotectant delivery and removal from murine insulinomas at vitrification-relevant concentrations
| Autor: | Athanassios Sambanis, Ying C. Song, Indra N. Mukherjee |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Cryopreservation
Cell Membrane Permeability Membrane permeability Cryoprotectant Dimethyl sulfoxide General Medicine Biology Article General Biochemistry Genetics and Molecular Biology Cell Line Propanediol Mice chemistry.chemical_compound Cryoprotective Agents Biochemistry chemistry Osmotic Pressure Biophysics Animals Osmotic pressure Insulinoma Vitrification General Agricultural and Biological Sciences Cytotoxicity |
| Zdroj: | Cryobiology. 55:10-18 |
| ISSN: | 0011-2240 |
| DOI: | 10.1016/j.cryobiol.2007.04.002 |
| Popis: | Development of optimal cryopreservation protocols requires delivery and removal of cryoprotective agents (CPAs) in such a way that negative osmotic and cytotoxic effects on cells are minimized. This is especially true for vitrification, where high CPA concentrations are employed. In this study, we report on the determination of cell membrane permeability parameters for water (L(p)) and solute (P(s)), and on the design and experimental verification of CPA addition and removal protocols at vitrification-relevant concentrations for a murine insulinoma cell line, betaTC-tet cells. Using membrane permeability values and osmotic tolerance limits, mathematical modeling and computer simulations were used to design CPA addition and removal protocols at high concentrations. The cytotoxic effects of CPAs were also evaluated. Cells were able to tolerate the addition and removal of 2.5M dimethyl sulfoxide (DMSO) and 2.5M 1,2 propanediol (PD) in single steps, but required multi-step addition and removal with 3.0M DMSO, 3.0M PD, and a vitrification-relevant concentration of 3.0M DMSO+3.0M PD. Cytotoxicity studies revealed that betaTC-tet cells were able to tolerate the presence of single component 6.0M DMSO and 6.0M PD and to a lesser extent 3.0M DMSO+3.0M PD. These results determine the time and concentration domain of CPA exposure that cells can tolerate and are essential for designing cryopreservation protocols for free cells as well as cells in engineered tissues. |
| Databáze: | OpenAIRE |
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