Development of Macromolecular Cryoprotectants for Cryopreservation of Cells.

Cryopreservation is a common way for long-term storage of therapeutical proteins, erythrocytes, and mammalian cells. For cryoprotection of these biosamples to keep their structural integrity and biological activities, it is essential to incorporate highly efficient cryoprotectants. Currently, permeable small molecular cryoprotectants such as glycerol and dimethyl sulfoxide dominate in cryostorage applications, but they are harmful to cells and human health. As acting in the extracellular space, membrane-impermeable macromolecular cryoprotectants, which exert remarkable membrane stabilization against cryo-injury and are easily removed post-thaw, are promising candidates with biocompatibility and feasibility. Water-soluble hydroxyl-containing polymers such as poly(vinyl alcohol) and polyol-based polymers are potent ice recrystallization inhibitors, while polyampholytes, polyzwitterions, and bio-inspired (glyco)polypeptides can significantly increase post-thaw recovery with reduced membrane damages. In this review, the synthetic macromolecular cryoprotectants are systematically summarized based on their synthesis routes, practical utilities, and cryoprotective mechanisms. It provides a valuable insight in development of highly efficient macromolecular cryoprotectants with valid ice recrystallization inhibition activity for highly efficient and safe cryopreservation of cells.

Self-Assembly of a C16M[Mn] Magnetic Surfactant in Water.

A magnetic surfactant, which combines the properties of a surfactant with magnetic responsiveness, shows great potential in biotechnology, separation, adsorption, and catalysis, especially in non-invasive manipulation through a magnetic field. However, a molecularly magnetic surfactant is usually paramagnetic for the amorphous and less ordered structures. In this work, magnetic surfactant 1-methyl-3-hexadecane-imidazolium [MnCl2Br] (C16M[Mn]) is reported to self-assemble in water. The C16M[Mn] magnetic surfactant self-assembles in water to form a lamellar hydrogel from 10 to 50 wt % at and below room temperature. The hydrogel changes from a gel to a sol at 30 °C, and the hexadecane chains in the hydrogel change from noncrystalline to crystalline at 0 °C. In the hydrogel state, the lamellar domain spacing is varied from 36 to 45 nm depending on the concentration and self-assembly temperature. After self-assembly, the magnetic susceptibility of the freeze-dried magnetic surfactant is increased. Most important is the fact that the freeze-dried sample at a high concentration (40-50 wt %) shows the highest magnetic susceptibility, which is related to the closer molecular packing and the more ordered structures. The self-assembly-induced increase in magnetic susceptibility provides a method for improving the magnetic properties of a magnetic surfactant.

Development of Icephilic ACTIVE Glycopeptides for Cryopreservation of Human Erythrocytes.

Ice formation and recrystallization exert severe impairments to cellular cryopreservation. In light of cell-damaging washing procedures in the current glycerol approach, many researches have been devoted to the development of biocompatible cryoprotectants for optimal bioprotection of human erythrocytes. Herein, we develop a novel ACTIVE glycopeptide of saccharide-grafted ε-poly(L-lysine), that can be credited with adsorption on membrane surfaces, cryopreservation with trehalose, and icephilicity for validity of human erythrocytes. Then, by Borch reductive amination or amidation, glucose, lactose, maltose, maltotriose, or trehalose was tethered to ε-polylysine. The synthesized ACTIVE glycopeptides with intrinsic icephilicity could localize on the membrane surface of human erythrocytes and improve cryopreservation with trehalose, so that remarkable post-thaw cryosurvival of human erythrocytes was achieved with a slight variation in cell morphology and functions. Human erythrocytes (∼50% hematocrit) in cryostores could maintain high cryosurvival above 74%, even after plunged in liquid nitrogen for 6 months. Analyses of differential scanning calorimetry, Raman spectroscopy, and dynamic ice shaping suggested that this cryopreservation protocol combined with the ACTIVE glycopeptide and trehalose could enhance the hydrogen bond network in nonfrozen solutions, resulting in inhibition of recrystallization and growth of ice. Therefore, the ACTIVE glycopeptide can be applied as a trehalose-associated "chaperone", providing a new way to serve as a candidate in glycerol-free human erythrocyte cryopreservation.

Up-regulated miR-374a-3p relieves lipopolysaccharides induced injury in CHON-001 cells via regulating Wingless-type MMTV integration site family member 5B.

BACKGROUND: Osteoarthritis (OA) is a frequent and incurable joint disease, inducing significant pain and seriously threatening to human health. It has been reported that microRNAs (miRNAs) play crucial roles on cancers and inflammatory diseases via cooperating with genes. However, the effect of miR-374a-3p/Wingless-type MMTV integration site family, member 5B (WNT5B) pair in OA remains to be explored. METHODS: GSE105027 and GSE55457 datasets were obtained to reveal the expression of miR-374a-3p and WNT5B in OA cartilages using log-scale. The OA cell model was established by lipopolysaccharides (LPS) stimulation in CHON-001 cells and the functional role of miR-374a-3p on OA was investigated by analyzing cell proliferation, cell apoptosis and the expression of apoptosis-related proteins (Bcl-2, Bax and Bim). Through bioinformatics prediction, WNT5B, the target gene of miR-374a-3p, was predicted and the association between miR-374a-3p and WNT5B was further explored by luciferase reporter assay. Functional experiments in vitro were conducted to assess whether WNT5B was involved in the regulation of miR-374a-3p to LPS-stimulated CHON-001. Finally, the expression of JNK/ERK/MAPK pathway-related proteins was detected to explore the underlying molecular mechanism. RESULTS: The data set showed that miR-374a-3p was decreased in OA cartilages and the consistent expressional pattern was observed in LPS-stimulated CHON-001 cells. Overexpression of miR-374a-3p significantly alleviated LPS-induced damage in CHON-001 cells, whereas miR-374a-3p inhibitor aggravated LPS-stimulated injury. Further experiments demonstrated that WNT5B was a target of miR-374a-3p and its expression was decreased by miR-374a-3p. WNT5B expression was increased in OA cartilages. Silencing WNT5B prevented CHON-001 cells from LPS-induced damage. Down-regulation of WNT5B strengthened the protective effect of miR-374a-3p on LPS-stimulated CHON-001 cells. Moreover, miR-374a-3p cooperated with WNT5B to affect cell behaviors of LPS-stimulated CHON-001 cells via mediating the JNK/ERK/MAPK pathway. CONCLUSION: These results indicated that overexpression of miR-374a-3p protects CHON-001 cells against LPS challenge by modulating WNT5B and inhibiting the JNK/ERK/MAPK pathway.

An injectable supramolecular hydrogel hybridized with silver nanoparticles for antibacterial application.

Silver nanoparticles (AgNPs) show long-lasting and broad-spectrum antibacterial activity. Herein, PEGylated AgNPs were prepared in situ by complexing AgNO3 with the random copolymer of poly(ethylene glycol) methyl ether methacrylate (PEGMA) and polyacrylic acid via electrostatic interaction followed by in situ reduction. AgNP hybrid supramolecular hydrogels were thus prepared through host-guest inclusion between PEGMA side chains and α-cyclodextrins in aqueous solution. The hydrogels were physically cross-linked by both pseudopolyrotaxane crystallization and AgNPs, which showed temperature responsiveness and self-healing properties. By hybridizing AgNPs, the hydrogels showed excellent antibacterial properties against S. aureus and E. coli bacteria as well as low cytotoxicity and have potential applications as injectable antibacterial materials.

Metal-containing and related polymers for biomedical applications.

A survey of the most recent progress in the biomedical applications of metal-containing polymers is given. Due to the unique optical, electrochemical, and magnetic properties, at least 30 different metal elements, most of them transition metals, are introduced into polymeric frameworks for interactions with biology-relevant substrates via various means. Inspired by the advance of metal-containing small molecular drugs and promoted by the great progress in polymer chemistry, metal-containing polymers have gained momentum during recent decades. According to their different applications, this review summarizes the following biomedical applications: (1) metal-containing polymers as drug delivery vehicles; (2) metal-containing polymeric drugs and biocides, including antimicrobial and antiviral agents, anticancer drugs, photodynamic therapy agents, radiotherapy agents and biocides; (3) metal-containing polymers as biosensors, and (4) metal-containing polymers in bioimaging.

Photocarrier transport and dynamics in mixed-phase BiFeO3 films.

We report a remarkable photoinduced relaxation process and its dependence of thickness and temperature in mixed-phase BiFeO3 films grown on (001) LaAlO3 substrates. When the films are illuminated by the light above the bandgap, their resistances are reduced with the increase of temperature. The photoinduced change of resistance reaches to the maximum of about 2.17 × 105% at 300 K. It is noted that the relaxation processes of the resistance are significantly different between T-like phase and T-R mixed phase due to structural strain, symmetry breaking and built-in electric field at the phase boundaries. These results provide more insights into intrinsic mechanisms of mixed-phase multiferroic materials and potential applications in all-oxide photoelectric devices.

Biocompatible Metal-Free Perovskite Membranes for Wearable X-ray Detectors.

Halide perovskites are emerging as promising materials for X-ray detection owing to their compatibility with flexible fabrication, cost-effective solution processing, and exceptional carrier transport behaviors. However, the challenge of removing lead from high-performing perovskites, crucial for wearable electronics, while retaining their superior performance, persists. Here, we present for the first time a highly sensitive and robust flexible X-ray detector utilizing a biocompatible, metal-free perovskite, MDABCO-NH4I3 (MDABCO = methyl-N'-diazabicyclo[2.2.2]octonium). This wearable X-ray detector, based on a MDABCO-NH4I3 thick membrane, exhibits remarkable properties including a large resistivity of 1.13 × 1011 Ω cm, a high mobility-lifetime product (µ-τ) of 1.64 × 10-4 cm2 V-1, and spin Seebeck effect coefficient of 1.9 nV K-1. We achieve a high sensitivity of 6521.6 ± 700 µC Gyair-1 cm-2 and a low detection limit of 77 nGyair s-1, ranking among the highest for biocompatible X-ray detectors. Additionally, the device exhibits effective X-ray imaging at a low dose rate of 1.87 µGyair s-1, which is approximately one-third of the dose rate used in regular medical diagnostics. Crucially, both the MDABCO-NH4I3 thick membrane and the device showcase excellent mechanical robustness. These attributes render the flexible MDABCO-NH4I3 thick membranes highly competitive for next-generation, high-performance, wearable X-ray detection applications.

Trends in women's representation as lead authors in high impact journals of lung cancer.

Background: Women's participation in the clinical oncology practice has increased over the past decade. There is a need to investigate whether women's participation in academia, as reflected by publication activity, increased over the time. This study aimed to investigate trends in female authorship in top journals of lung cancer in the past ten years. Methods: In this cross-sectional study of all original research and review articles published in lung cancer journals including New England Journal of Medicine, Lancet journals, JAMA journals, Journal of Clinical Oncology, Annals of Oncology, Cancer Discovery, Journal of Thoracic Oncology, and Translational Lung Cancer Research (TLCR) between 2012 and 2021, the sex composition of lead authors were analyzed. The sex of the author was confirmed by internet searching for photographs, biographies, and gender-specific pronouns from journal or personal websites. The time-trend of female authorship was determined using Join-Point Regression (JPR) analysis. Results: A total of 3,625 first authors and 3,612 corresponding authors were identified in the journals during the years studied. The sex of the author was revealed for 98.5%. Among 3,625 first authors with the sex being revealed, 1,224 (33.7%) were women. The proportion of female first authors increased remarkably from 29.4% in 2012 to 39.8% in 2021. The annual percentage change (APC) in female first authorship took place in 2019 [APC for 2019-2021, 37.03, 95% confidence interval (CI): 18.0-59.1, P=0.003]. The proportion of first authors in TLCR increased from 25.9% in 2012 to 42.8% in 2021 and showed the greatest rise in female first authorship. There were significant discrepancies in the female first authorship across the journals and regions. Among the 3,612 corresponding authors whose sex were determined, 884 (24.5%) were female. There is no significant increasing trend in female corresponding authorship. Conclusions: The sex gap in the first authorship of lung cancer research articles has improved markedly in the recent years, however, sex imparities persist especially in corresponding authorship. There is an urgent need to proactively support and promote women in taking the leadership roles, thereby increasing their contributions to and influence on the development or advancement for future healthcare policies and practices.

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

Cryopreservation of red blood cells (RBCs) is imperative for transfusion therapy, while cryoprotectants are essential to protect RBCs from cryoinjury under freezing temperatures. Trehalose has been considered as a biocompatible cryoprotectant that naturally accumulates in organisms to tolerate anhydrobiosis and cryobiosis. Herein, we report a feasible protocol that enables glycerol-free cryopreservation of human RBCs by integration of the synthesized trehalose lipids and dissociative trehalose through ice tuning and membrane stabilization. Typically, in comparison with sucrose monolaurate or trehalose only, trehalose monolaurate was able to protect cell membranes against freeze stress, achieving 96.9 ± 2.0% cryosurvival after incubation and cryopreservation of human RBCs with 0.8 M trehalose. Moreover, there were slight changes in cell morphology and cell functions. It was further confirmed by isothermal titration calorimetry and osmotic fragility tests that the moderate membrane-binding activity of trehalose lipids exerted cell stabilization for high cryosurvival. The aforementioned study is likely to provide an alternative way for glycerol-free cryopreservation of human RBCs and other types of cells.

A Dynamic Membrane-Active Glycopeptide for Enhanced Protection of Human Red Blood Cells against Freeze-Stress.

Intracellular delivery of freezing-tolerant trehalose is crucial for cryopreservation of red blood cells (RBCs) and previous strategies based on membrane-disruptive activity usually generate severe hemolysis. Herein, a dynamic membrane-active glycopeptide is developed by grafting with 25% maltotriose and 50% p-benzyl alcohol for the first time to effectively facilitate entry of membrane-impermeable trehalose in human RBCs with low hemolysis. Results of the mechanism acting on cell membranes suggest that reversible adsorption of such benzyl alcohol-grafted glycopeptide on cell surfaces upon weak perturbation with phospholipids and dynamic transition toward membrane stabilization are essential for keeping cellular biofunctions. Furthermore, the functionalized glycopeptide is indicative of typical α-helical/ß-sheet structure-driven regulations of ice crystals during freeze-thaw, thereby strongly promoting efficient cryopreservation. Such all-in-one glycopeptide enables achieving both high cell recovery post-thaw >85% and exceptional cryosurvival >95% in direct freezing protocols. The rationally designed benzyl alcohol-modified glycopeptide permits the development of a competent platform with high generality for protection of blood cells against freeze-stress.

High-fiber diet ameliorates gut microbiota, serum metabolism and emotional mood in type 2 diabetes patients.

Previous studies have demonstrated that patients with type 2 diabetes mellitus (T2DM) often had the problems of fecal microbiota dysbiosis, and were usually accompanied with psychiatric comorbidities (such as depression and anxiety). Here, we conducted a randomized clinical study to analyze the changes in gut microbiota, serum metabolism and emotional mood of patients with T2DM after consumption of a high-fiber diet. The glucose homeostasis of participants with T2DM was improved by the high-fiber diet, and the serum metabolome, systemic inflammation and psychiatric comorbidities were also altered. The increased abundances of Lactobacillus, Bifidobacterium and Akkermansias revealed that the proportions of beneficial gut microbes were enriched by the high-fiber diet, while the abundances of Desulfovibrio, Klebsiella and other opportunistic pathogens were decreased. Therefore, the current study demonstrated that the intestinal microbiota alterations which were influenced by the high-fiber diet could improve the serum metabolism and emotional mood of patients with T2DM.

Cobaltocenium-containing block copolymers: ring-opening metathesis polymerization, self-assembly and precursors for template synthesis of inorganic nanoparticles.

Side-chain cobaltocenium-containing block copolymers are prepared by ring-opening metathesis polymerization (ROMP). These block copolymers include one cobaltocenium-containing block, with the second block being either a nonmetal-containing segment or a cobaltocenium-containing segment with different counterions. These block copolymers are self-assembled into spherical core/shell micelles in solutions. A template strategy is used to prepare cobalt (II or III)-containing nanoparticles by treating the self-assembled micelles via UV/ozonolysis and pyrolysis. Characterization by X-ray photon spectroscopy and X-ray diffraction indicates that these nanoparticles consist of different oxidants of cobalt, depending on the chemical compositions of block copolymers.

Sulfonium-Containing Glycopolypeptides Tethering Trehalose for Protein Stabilization.

Protection of proteins is of great significance since external stimuli can cause denaturation due to aggregation. Herein, a series of well-defined cationic trehalose-glycopolypeptides, poly(ethylene glycol)-b-(l-methionine-g-trehalose)s (PEG45-b-(Met-g-Tre)x), were synthesized for stabilization of a model protein, glucose oxidase (GOx), against lyophilization and heating. The isothermal titration calorimetry results revealed that polyionic complexes (PICs) were formed between the synthesized trehalose-glycopolypeptides and GOx via electrostatic interactions. These PICs allowed GOx to maintain an approximately 80% enzymatic activity compared with native GOx after 6× lyophilization. Meanwhile, PEG45-b-(Met-g-Tre)x also provided a positive effect on the protection of GOx upon heating at 60 °C. Results of transmission electron microscopy suggested that the trehalose-glycopolypeptides could prevent protein aggregation, thereby maintaining the bioactive function of GOx. In brief, it provided a synthesis strategy for the precise preparation of trehalose-glycopolypeptides, as well as a suitable method for stabilization of proteins.

Achieving high intracellular trehalose in hRBCs by reversible membrane perturbation of maltopyranosides with synergistic membrane protection of macromolecular protectants.

Trehalose is considered as a biocompatible cryoprotectant for solvent-free cryopreservation of cells, but the difficulty of the current trehalose delivery platforms to human red blood cells (hRBCs) limits its wide applications. Due to cell injuries caused by incubation at 37 °C and low intracellular loading efficiency, development of novel methods to facilitate trehalose entry in hRBCs is essential. Herein, a reversible membrane perturbation and synergistic membrane stabilization system based on maltopyranosides and macromolecular protectants was constructed, demonstrating the ability of efficient trehalose loading in hRBCs at 4 °C. Results of confocal laser scanning microscopy exhibited that the intracellular loading with the assistance of maltopyranosides was a reversible process, while the membrane protective effect of macromolecular protectants on trehalose loading in hRBCs was necessary. It was suggested that introduction of 30 mM poly(vinyl pyrrolidone) 8000 combined with 1 mM dodecyl-ß-D-maltopyranoside and 0.8 M trehalose could increase the intracellular trehalose to 84.0 ± 11.3 mM in hRBCs, whereas poly(ethylene glycol), dextran, human serum albumin or hydroxyethyl starch had a weak effect. All the macromolecular protectants could promote the cryosurvival of hRBCs, exhibiting membrane stabilization, and incubation and followed by cryopreservation did not change the basic functions and normal morphology of hRBCs substantially. This study provided an alternative strategy for glycerol-free cryopreservation of cells and the delivery of membrane-impermeable cargos.

NonFreezable Preservation of Human Red Blood Cells at -8 °C.

Red blood cell (RBC) preservation is very important in human health. The RBCs are usually preserved at 4 ± 2 °C without freezing or at a very low temperature (-80 °C or liquid nitrogen) with deep freezing. Herein, non freezable preservation of RBCs at a subzero temperature is reported to prolong the preservation time compared with that at 4 ± 2 °C. By adding glycerol and poly(ethylene glycol) (PEG) (average number molecular weight 400, PEG-400) into the preservation solution, the freezing point is decreased and the hemolysis is kept low. The cell metabolism of stored RBCs at -8 °C is reduced, and the shelf life of RBCs extends up to at least 70 days. At the end of preservation, the pH decreases a little bit to demonstrate the low metabolic rate of RBCs stored at subzero temperatures. After quick washing, the RBC survival rate is ca. 95%. The adenosine triphosphate, 2,3-diphosphoglycerate, and cell deformation ability of the washed RBCs are maintained at a high level, while the malondialdehyde is relatively low, which verifies the high quality of RBCs stored at this condition.

[Effect of Anti-Oxidative of Ethyl Pyruvate and Taurine on the Red Blood Cell Storage at 4 ℃].

OBJECTIVE: To investigate the anti-oxidative effect of ethyl pyruvate (EP) and taurine (TAU) on the quality of red blood cells stored at 4±2 ℃, hemolysis, energy metabolism and lipid peroxidation of the red blood cells in the preservation solution were studied at different intervals. METHODS: At 4±2 ℃, the deleukocyte red blood cells were stored in the citrate-phosphate-dextrosesaline-adenine-1 (CPDA-1) preservation (control group), preservation solution with EP (EP-AS), and TAU (TAU-AS) for long-term preservation. The enzyme-linked immunoassay and automatic blood cell analyzer were used to detect hemolysis and erythrocyte parameters. Adenine nucleoside triphosphate (ATP), glycerol 2,3-diphosphate (2,3-DPG) and malondialdehyde (MDA) kits were used to test the ATP, 2,3-DPG and MDA concentration. RESULTS: During the preservation, the rate of red blood cell hemolysis in EP-AS and TAU-AS groups were significantly lower than that in CPDA-1 group (P<0.01). The MCV of EP-AS group was increased with the preservation time (r=0.71), while the MCV of the TAU-AS group was significantly lower than that in the other two groups (P<0.05). The concentration of ATP and MDA in EP-AS and TAU-AS groups were significantly higher than that in CPDA-1 group at the 14th day (P<0.01). The concentrations of 2,3-DPG in the EP-AS and TAU-AS groups were significantly higher than that in the CPDA-1 group from the 7th day (P<0.01). CONCLUSION: EP and TAU can significantly reduce the red blood cell hemolysis rate, inhibit the lipid peroxidation level of red blood cells, and improve the energy metabolism of red blood cells during storage. The mechanism of EP and TAU may be related to their antioxidation and membrane protection effect, so as to improve the red blood cell quality and extend the preservation time.

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

Cryopreservation of human erythrocytes via suitable cryoprotectants is essential for transfusion during emergencies, but the conventional glycerolization method requires a tedious thawing-deglycerolization process. Alternatively, trehalose, a nonreducing disaccharide, has gained much attention as a biocompatible cryoprotectant due to its nature in living organisms capable of surviving extreme cold and desiccation. In this work, cryopreservation of human erythrocytes was realized through high intracellular trehalose enhanced by benzyl alcohol at 4 °C with membrane stabilization of maltotriose-grafted ε-poly(L-lysine). Intracellular trehalose could reach 94.2 ± 12.1 mM with slight impacts on morphology and cell functions, and the post-storage cryosurvival of human erythrocytes could achieve 96.2 ± 3.4% via membrane protection by the glycopeptide. It has been demonstrated that the functional glycopeptide performed as an extracellular cryoprotectant accompanied by high intracellular trehalose for synergistic cryopreservation of human erythrocytes in the biocompatible glycerol-free conditions. This two-step approach involving augmentation of intracellular trehalose at a hypothermic temperature and membrane stabilization of the functional glycopeptide could be an alternative way for human cell cryopreservation.

Facilitating trehalose entry into hRBCs at 4 °C by alkylated ε-poly(L-lysine) for glycerol-free cryopreservation.

Currently, glycerol is a conventional cryoprotectant of human red blood cells (hRBCs), but the time-consuming thawing and deglycerolization processes are essential before transfusion. Much of the research up to now has been conducted on the delivery of impermeable trehalose to hRBCs at 37 °C, but the cryoprotective effect of trehalose and deterioration of cells still remain challenging. Encouraged by the interaction of hydrophobic or cationic groups on cell membranes and osmotic stabilization, herein, we propose a novel cryopreservation system to facilitate trehalose entry into hRBCs at 4 °C and pH 7.4. High intracellular trehalose contents and cryosurvival of hRBCs were achieved with small function variations via the assistance of self-assembled nanoparticles of alkylated ε-poly(L-lysine) (ε-PL) along with poly(vinyl pyrrolidone) (PVP). The effect of amphipathic alkylated ε-PL with various alkyl chains and grafting ratios on membrane perturbation with protection of PVP was systematically investigated. Overall, by the combination of alkylated ε-PL and PVP, the intracellular trehalose could be enhanced to 109.7 ± 6.1 mM and subsequently hRBC cryosurvival reached 91.7 ± 5.5%, significantly higher than those containing trehalose only, 11.9 ± 1.1 mM and 50.0 ± 2.1%, respectively. It was observed that the biocompatible trehalose-loading system could benefit glycerol-free cryopreservation of hRBCs and also provide a feasible way for impermeable biomacromolecule delivery.

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

As a nonreducing disaccharide, trehalose can be used as a biocompatible cryoprotectant for solvent-free cell cryopreservation, but the membrane-impermeability limits its cryoprotective efficiency. Herein, a series of aromatic monoamines with a 1-4 methylene spacer were grafted onto γ-poly(glutamic acid) (γ-PGA) for promoting intracellular trehalose uptake in human red blood cells (hRBCs) via membrane perturbation. The self-assembled nanoparticles of the obtained amphiphilic γ-PGA could be adsorbed on the cell membrane by the hydrophobic interaction to disturb the lipid arrangement and increase the membrane permeability of trehalose under hypertonic conditions. Results suggested that the intracellular trehalose could be enhanced progressively with the methylene spacer length, significantly increasing to 75.1 ± 0.7 mM by incubating hRBCs in 0.8 M trehalose containing phenylbutylamine-grafted γ-PGA at 4 °C for 24 h. Meanwhile, the other three polymers exhibited membrane stabilization in addition to improved intracellular trehalose, maintaining the membrane integrity during cryopreservation to achieve high cryosurvival. Molecular dynamics simulation further confirmed that defects could be formed by interaction of the above four amphiphilic polymers on the modeled phospholipid bilayer. It was believed that glycerol-free cryopreservation of human cells could be realized by using trehalose as the biocompatible cryoprotectant, and membrane stabilization can be a compensatory approach to membrane perturbation during impermeable biomolecule delivery.