Cryopreservation of human erythrocytes through high intracellular trehalose with membrane stabilization of maltotriose-grafted ε-poly(l-lysine)

Abstract

Cryopreservation of human erythrocytes via suitable cryoprotectants is essential for transfusion at emergency, but the conventional glycerolization method requires a tedious thawing-deglycerolization process. Alternatively, trehalose, a nonreducing disaccharide, has gained...

Integration of Trehalose Lipids with Dissociative Trehalose Enables Cryopreservation of Human RBCs

“…After incubation and cryopreservation, the samples were centrifuged at 1000 g for 7 min, and the absorbance ( A ) of the supernatant was measured via a spectrophotometer (Tecan, Switzerland) at 540 nm. The hemolysis was calculated according to eq − hemolysis 0.25em ( % ) = A sample A positive × 100 % …”

“…The intracellular trehalose content in hRBCs was determined by the anthrone colorimetry method using a trehalose kit (Beijing Solarbio, China) after incubation. − For samples containing trehalose, after incubation at 4 °C for 20 h, the suspension was centrifuged at 1000 g for 7 min with the supernatant discarded and repeatedly washing three times to remove residual extracellular trehalose. The washed hRBCs were diluted 10 times using isotonic PBS solution, and the red blood cell count (RBC) and mean cell volume (MCV) of the samples were measured using an automated hematology analyzer (Bowlinman BM830, Beijing, China).…”

“…Along with the aforementioned hypotheses, it has been demonstrated that trehalose offers optimal cryoprotection when dispersed on both sides of the cell membrane . Consequently, there have been considerable works exploring strategies to promote intracellular delivery of nonpermeable trehalose. − For erythrocytes without the ability of endocytosis, different methods such as osmotic stress, electroporation, polymers, and peptides have been employed to facilitate intracellular trehalose uptake. For instance, an electroporation technology was attempted to introduce trehalose into hRBCs by allowing electrical pulses to pass through the cell membrane .…”

Membrane stabilization versus perturbation by aromatic monoamine-modified γ-PGA for cryopreservation of human RBCs with high intracellular trehalose

“…Then, the hRBC suspension was immersed in liquid nitrogen and frozen at a cooling rate of approximately 140 1C min À1 . 48 After being stored in liquid nitrogen for more than 2 h, the frozen samples were thawed in a 37 1C water bath. The cryosurvival was calculated according to the eqn (3): 17,28,29 Cryosurvival % ð Þ ¼ Cell count after freeze-thaw Cell count after incubation  100…”

Sulfonium-Containing Glycopolypeptides Tethering Trehalose for Protein Stabilization

“…Inspired by glycoproteins, biomimetic glycopolymers with carbohydrates conjugating to the polymeric backbone could realize important physiological functions . Alternatively, glycopolypeptides could be used for protection of biological units due to their well-defined structures, which were dominant in morphological control compared with natural biomacromolecules. , In our previous work, ε-poly­( l -lysine)­s with carboxylated trehalose pendant were adopted to protect red blood cells from cryo-injury, exhibiting excellent performance of membrane stabilization. − Recently, poly­( l -alanine- co - l -lysine)- g -trehalose was also prepared as a biomimetic cryoprotectant for stem cells . However, so far, there are few reports on trehalose-containing glycopolypeptides for protein preservation.…”