A Paradigm Shift in Cryopreservation: Molecular-Based Advances to Improve Outcome

Assessment of the Impact of Post-Thaw Stress Pathway Modulation on Cell Recovery following Cryopreservation in a Hematopoietic Progenitor Cell Model

“…Furthermore, it was found that this molecular control strategy could reduce the need for traditional cryoprotective agents (CPA) such as DMSO [ 37 , 38 ]. This discovery resulted in a paradigm shift in the cryopreservation sciences from a primarily chemo-osmometric (ice control) approach to that of an integrated approach combining molecular biological control with ice control [ 24 , 39 , 40 , 41 , 42 ].…”

“…While many researchers remain unaware, today, this shift is mainstream and serves as the foundational basis of today’s commercial intracellular-like cryopreservation media such as CryoStor ® and Unisol™ [ 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 ]. Functionally, through their multi component formulation, intracellular-like solutions buffer the environment a cell is exposed to during the freeze−thaw process, modulating stress response activation, thereby reducing CIDOCD [ 23 , 25 , 41 , 42 , 50 ]. Numerous reports have detailed the benefits of using intracellular-like solutions in comparison to traditional extracellular-like cell culture media with DMSO [ 22 , 46 , 47 , 51 , 52 , 53 , 54 , 55 , 56 ].…”

“…Today, 100’s of studies have demonstrated >50% improvement in cell recovery and function through the use of intracellular-like solutions in an equal number of unique cell and tissue systems, ranging from stem cells to CAR-T cells to engineered tissues. While taking over a decade for this shift to become an integral part of the cryopreservation sciences, intracellular-like solutions are now the gold standard for modern cryopreservation [ 41 , 42 , 44 , 45 , 54 , 55 , 57 ].…”

“…While the integration of cellular−molecular control and the introduction of intracellular-like cryopreservation solutions has had a transformative impact providing for significant increases in the recovery, repopulation, function, and structural integrity of cells and tissues (native and engineered) post-thaw, several challenges remain to enable an even greater impact. These include (1) continued cell loss (ranging from 10 to >50% depending on the cell type), (2) compromised function in complex cells and tissues, (3) spontaneous differentiation in native stem cell populations, (4) dependence on DMSO, and (5) lack of/limited utility of equipment used in the freezing and thawing processes [ 1 , 2 , 41 , 54 , 55 , 56 , 57 , 58 ]. Furthermore, recognition of the critical role cryopreservation plays in the success of the field of regenerative medicine remains a hurdle.…”

“…Furthermore, recognition of the critical role cryopreservation plays in the success of the field of regenerative medicine remains a hurdle. In an effort to address these challenges and improve the outcome, today’s researchers are focused on developing new CPAs [ 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 ], targeted apoptotic control [ 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 ] during cryopreservation and new devices [ 69 , 70 , 71 ]. On the discovery science side, groups continue to investigate the pathways activated during cryopreservation, as well as during the recovery phase [ 72 , 73 , 74 , 75 , 76 , 77 ].…”