Cryopreservation of bovine somatic cells using antifreeze polyamino-acid (carboxylated poly- l -lysine)

Pan

Journal of Nanomaterials

et al. 2021

Cryopreservation has been a key technology in medical science, food preservation, and many other fields. In a freezing process, the formation of ice crystals may cause significant damage to the frozen samples. In order to reduce the damage, many cryoprotectants (CPAs) have been developed and added in cryopreservation processes for reduced ice volume, decreased ice size, proper ice shaping, and cell protection. According to the material characteristics, the CPAs are either impermeable (i.e., antifreeze protein, polyampholytes, and polyvinyl alcohol) or permeable (i.e., dimethyl sulfoxide, proline, and glycerol) to cell membranes. The typical CPAs are introduced in this work with their material characteristics, antifreeze mechanisms, and applications. Antifreeze mechanisms for different CPAs involve molecular adsorption on the ice surface, hydrogen bonding to ice, bending the ice surface, lowering the freezing point, inhibiting ice recrystallization, protecting cell membranes, reducing dehydration of cells, and breaking hydrogen bonds among ice crystals to reduce the size of ice crystals. In practice, different CPAs can be used together with their cryopreservation properties functioning synergetically. This study reviews the recent applications of CPAs in food, biology and medicine, and agriculture. Future works for CPAs are suggested in improving efficiency, revealing mechanisms, broadening application, and finding new CPAs.

Section: Polyampholytementioning

confidence: 99%

A Review of the Material Characteristics, Antifreeze Mechanisms, and Applications of Cryoprotectants (CPAs)

Pan

Journal of Nanomaterials

et al. 2021

“…Recently, carboxylated ε-poly-L-lysine (CPLL) has been utilized, a bioinspired biocompatible material that has been used as a food and cell preservative as well as for tissue engineering and drug delivery in the biomedical applications because of its reduced toxicity and cryoprotective properties [17,18]. In addition, CPLL has also been reported for use as an additional CPA in the cryopreservation of numerous cell types such as the red blood cells [19], natural killer cells [17], stem cells [20], and bovine somatic cells [21]. Furthermore, CPLL showed a higher cryoprotective property effect on pig embryos [22] and Nili-Ravi buffalo bull sperm [23].…”

Section: Introductionmentioning

confidence: 99%

Carboxylated ε-Poly-L-Lysine Supplementation of the Freezing Extender Improves the Post-Thawing Boar Sperm Quality

Zhang¹,

Zhu²

2022

Animals

Frozen boar sperm is used on a minimal scale in consequence of the cryo-injuries induced by biochemical and physical modifications during the freezing and thawing processes. The present study investigates whether the addition of carboxylated ε-poly-L-lysine (CPLL) to the freezing medium could improve post-thaw boar sperm quality or not. Boar sperm was diluted with freezing medium contained different doses of carboxylated ε-poly-L-lysine (0, 0.125%, 0.25%, 0.5%, and 1%; v/v). The motility patterns, membrane integrity, acrosome integrity, mitochondrial membrane potential, NADH-CoQ activity, ATP level, malondialdehyde (MDA) level, and antioxidant defense system, as well as apoptosis in post-thaw boar sperm, were measured. It was observed that 0.25% CPLL treatment significantly improved the post-thaw boar sperm total motility, progressive motility, straight-linear velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), linearity (LIN), straightness (STR), membrane integrity, and acrosome integrity. Interestingly, the addition of CPLL also significantly increased the post-thaw sperm mitochondrial membrane potential, NADH-CoQ activity, and ATP level. Moreover, post-thaw boar sperm catalase (CAT) activity, glutathione peroxidase (GPx) activity, and superoxide dismutase (SOD) activity were increased with the addition of CPLL from 0.125% to 0.5% concentration levels. Furthermore, reduction of post-thaw sperm MDA level and apoptosis in 0.25% CPLL treatment was also observed. Those observations suggested that the addition of 0.25% CPLL to the freezing medium increased post-thaw boar sperm quality by protecting sperm mitochondrial function and antioxidant defense system. These findings provided novel insights that CPLL can be used as an efficient cryoprotectant to improve the post-thaw boar sperm quality during cryopreservation.

“…It has been used as a food and cell preservative as well as for other biomedical applications (e.g., tissue engineering and drug delivery) . Recently, the cryoprotective property of CPLL has been reported for numerous cell types such as red blood cells and stem cells . Carboxylated PLL acts in a way similar to antifreeze proteins (AFPs); however, the exact mechanism of action is still unknown .…”

mentioning

confidence: 99%

“…Therefore, ice recrystallization forms a matrix of large ice crystals, causing more damage to cells as a result of membrane rapture and cell dehydration . In addition, the ice crystals in CPLL are more hexagonal or bipyramid in shape since the carboxyl group binds to crystals . This hexagonal crystal shape reduces the damage to cell membrane, in contrast to round and flat ice crystals observed when cooling pure water .…”

mentioning

confidence: 99%

“…In addition, the ice crystals in CPLL are more hexagonal or bipyramid in shape since the carboxyl group binds to crystals . This hexagonal crystal shape reduces the damage to cell membrane, in contrast to round and flat ice crystals observed when cooling pure water . Furthermore, the CPLL shows high adsorption to cell membrane during cooling, which also contributes to the protection of cell membrane from outside and maintaining membrane integrity .…”

mentioning

confidence: 99%

See 1 more Smart Citation

Bioinspired Preservation of Natural Killer Cells for Cancer Immunotherapy

et al. 2019

The ability to cryopreserve natural killer (NK) cells has a significant potential in modern cancer immunotherapy. Current cryopreservation protocols cause deterioration in NK cell viability and functionality. This work reports the preservation of human cytokine‐activated NK cell viability and function following cryopreservation using a cocktail of biocompatible bioinspired cryoprotectants (i.e., dextran and carboxylated ε‐poly‐L‐lysine). Results demonstrate that the recovered NK cells after cryopreservation and rewarming maintain their viability immediately after thawing at a comparable level to control (dimethyl sulfoxide‐based cryopreservation). Although, their viability drops in the first day in culture compared to controls, the cells grow back to a comparable level to controls after 1 week in culture. In addition, the anti‐tumor functional activity of recovered NK cells demonstrates higher cytotoxic potency against leukemia cells compared to control. This approach presents a new direction for NK cell preservation, focusing on function and potentially enabling storage and distribution for cancer immunotherapy.