Rui Li

This investigation traces the cryobiological challenges of a multi-stage hiPSC-derived differentiation and manufacturing process. Low-temperature Raman spectroscopy and a series of cell assays revealed that cell cryobiology changed as the cells differentiated from hiPSCs via neural crest cells to sensory neurons. We identified cell dissociation and incubation before freezing as two pivotal non-cryopreserved steps influencing the success of cryopreservation. Our study correlated membrane… Show more

This investigation traces the cryobiological challenges of a multi-stage hiPSC-derived differentiation and manufacturing process. Low-temperature Raman spectroscopy and a series of cell assays revealed that cell cryobiology changed as the cells differentiated from hiPSCs via neural crest cells to sensory neurons. We identified cell dissociation and incubation before freezing as two pivotal non-cryopreserved steps influencing the success of cryopreservation. Our study correlated membrane fluidity and cell size respectively with stage-specific sensitivity to nucleation temperature and cooling rate, which may be used to inform stage-specific cryopreservation designs to mitigate these sensitivities. This open-access article scrutinizes this manufacturing science from the combined perspective of cell development and cryobiology. Show less

Natural deep eutectic systems (NADES) are emerging as potential cryoprotective agents (CPA) for cell preservation. In this investigation, we develop an optimized CPA formulation using trehalose‐glycerol NADES (T:G) diluted in Normosol‐R and supplemented with isoleucine. The study also indicates that thermophysical properties of CPA solutions alone cannot predict optimal cell survival, suggesting that stabilization of biological structures by CPAs may play a role in successful cryopreservation.

This open access article highlights a DMSO-free approach to cryopreserve iPS cells in the form of multicellular aggregates with little loss in cell quantity and function; a strategy to mitigate cryopreserved cells' vulnerability to undercooling (uncontrolled ice nucleation/seeding at low temperatures); low-temperature Raman spectroscopy evidence of synergistic effect of sugar, sugar alcohol, amino acid and P188.

This article is a timely review of the state of cell-based immunotherapies. It addresses the key issues of interest that affect the quality of the cryopreserved cellular products, the current standard of practice that clinical studies have adopted, as well as the emerging technologies that aim to improve the preservation of these cellular therapeutics.

This article is a part of Interfaces and Biology 1: Mechanobiology and Cryobiology special issue.

Physiologically relevant in vitro 3D human organ model featuring 3D endothelium, 95% interaction area between 3D ECM and microfluidic channel, chemotaxis gradient for study of interstitial cancer cell migration, high-multipotency 3D stem cell microenvironment.

This study presented the significant difference between multicellular aggregates and single cells of the same cell type in response to freezing, the benefit of controlled-rate freezing with defined ice nucleation temperature, and an effective freeze-thaw protocol preserving the performance of human iPS cells at a level similar to standard passaging.