DMSO-free cryopreservation of chondrocytes based on zwitterionic molecule and polymers

Abstract

Cartilage tissue engineering highly relies on the ability to store and transport chondrocytes in order to be clinically successful. Cryopreservation is a most reliable technology for chondrocyte storage, but it suffers from the intrinsic toxicity of current state-of-the-art cryoprotectant, dimethyl sulfoxide (DMSO). In this work, we used the first fully zwitterionic compound-based approach for effective chondrocyte cryopreservation. A zwitterionic molecule combined with zwitterionic polymers could balance intra/ extracellular osmotic stress and prevent ice formation, which were the keys of successful cryopreservation. Moreover, this zwitterionic combination showed noncytotoxicity due to its high biocompatibility, superior to cytotoxic DMSO. On the basis of these performances, chondrocytes could be well cryopreserved (∼90% post-thaw survival efficiency) for a long time without any addition of DMSO, and the recovered cells could maintain their normal functionalities. In view of the association between polymer molecular weight and cryopreservation efficacy, further mechanism of cryoprotection provided by zwitterionic molecule/polymer was proposed. This work opens a new window of opportunity for DMSO-free cryopreservation using biocompatible zwitterionic materials.

Dimethyl Sulfoxide-Free Cryopreservation of Human Umbilical Cord Mesenchymal Stem Cells Based on Zwitterionic Betaine and Electroporation

“…However, continually increasing the betaine concentration in the cryo-medium did not improve the post thaw cell survival rate. There may exist a maximum of intracellular betaine absorbed by UCMSCs, which is likely related to the transmembrane transport manner of betaine and the expression of transporter on UCMSCs [ 32 ]. According to the result, this maximum of intracellular betaine could only provide limited protection during UCMSCs cryopreservation.…”

“…This potentials supported its selection as a novel cell cryopreservation CPA. However, for specific cell types like chondrocytes, stem cells, and dendritic cells, betaine alone cannot work well because of insufficient intracellular cryoprotections [ 32 ].…”

Macromolecular cryoprotectants for the preservation of mammalian cell culture: lessons from crowding, overview and perspectives

“…Thus, the combination of betaine and PSBMA could provide intracellular and extracellular cryopreservation. 60 Miermont et al observed that increasing osmotic compression decreases protein mobility, resulting in the cessation of several cellular processes. The rate of decrease in cellular processes was exponential when the osmotic compression was on the verge of stalling.…”

“…Chondrocytes were able to attach to substrates with a regular morphology, as opposed to cells cryopreserved in DMSO, which floated as the normal chondrocyte population was also significantly altered. 60 Besides this, Yang et al reported the use of natural biocompatible osmoprotectants in protecting cells from osmotic injury and serve as an ideal CPA. The biocompatible osmoprotectants -proline, glycine, and taurine -were assessed for their cytotoxicity.…”

“…86,87,[114][115][116] Additionally, zwitterionic polymers like PSBMA and PCBMA have demonstrated the ability to cryoprotect mammalian cells in combination with zwitterionic molecules such as betaine. 60,89 Furthermore, the application of polymeric hydrogels in cryopreservation of mammalian cell cultures has been described as follows.…”