Improved Hypothermic Preservation of Human Renal Cells Through Suppression of Both Apoptosis and Necrosis

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

“…Although, our data suggested that the presence of 1% MC, as the sole CPA, with DMSO and without DMSO, that is, even the presence of 2% DMSO was signifi cant and this extremely low concentration was able to cryoprotect P1 ASCs. cell viability for in vivo transplantation applications [58]. For example, Hollister et al [59] demonstrated that apoptosis is a signifi cant component of cell death in a human prostate cancer cell line (PC3) exposed to cryopreservation temperatures.…”

“…Since, apoptosis is implicated in the delayed onset of cell death and is accompanied by decline in attachment rate and long-time survival, it is important to perform viability assessment at different time points of post-thaw cultured cells. However, assessments at 24 h post-thaw allow for a manifestation of most components of the cell stress cascades and encompass the peak of apoptotic (8-12 h) and necrotic (4-8 h) activities [56][57][58]. Hence we have performed fl ow cytometry analysis of apoptosis and necrosis of ASCs, after a 24-h post-thaw incubation, for all the cryopreservation media investigated.…”

“…Recent evidence suggests that the freeze/thawing process induces extensive early apoptosis stress activation pathways, which can lead to a time-dependent decline in viability and function at culture temperatures [55][56][57][58][59][60][61]. Therefore, consideration must be given to the adoption of methods that simultaneously detect early apoptotic cells along with necrotic cells for a more accurate assessment of post-thaw in this study [62][63][64] and hence ensure the total solidifi cation (and long-term stability) of the frozen sample.…”

Low temperature cell pausing: an alternative short-term preservation method for use in cell therapies including stem cell applications

“…Osmotic imbalances can injure cells and programmed cell death may be induced upon thawing (Mathew et al 2002). Further to this, dimethyl sulfoxide (DMSO), the most frequently used CPA, is considered toxic to cells at ambient temperatures (Hunt 2011) and may induce cell differentiation (Santos et al 2003).…”

“…These cold storage media aim to support cell metabolism and inhibit post-storage necrosis and activation of apoptosis in response to cold temperatures (Hope et al 2011). HTS-FRS has been particularly successful for the storage of cells at 4°C (Mathew et al 2002;Mathew et al 2004). Comprised of ionic components such as Na + and Cl -, pH buffers, energy substrates and osmotic and oncotic stabilizers, the ionic makeup of HTS aims to balance cellular ion concentrations which are altered upon hypothermia and nutrient deprivation (Mathew et al 2004).…”