Sonoporation enables high-throughput loading of trehalose into red blood cells

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

“…[11][12][13] Generally, trehalose was delivered to cells by physical methods or with the assistance of hydrophobically modified polymers via membrane-disruptive activity, and these processes were usually carried out at 37 1C, above the phase transition temperatures of membrane phospholipids. 15,16,[18][19][20][21][22]28 In order to facilitate the entry of trehalose into human erythrocytes at 4 1C, in this work, human erythrocytes were first incubated with hypertonic extracellular trehalose containing the penetration enhancer benzyl alcohol to obtain high intracellular trehalose. Moreover, the functional glycopeptide, PL-g-M, was introduced and its membrane stabilizing activity for human erythrocytes was systematically investigated during incubation and freeze-thaw.…”

“…[11][12][13] In light of the difficulty for trehalose to either self-synthesize in mammalian cells or penetrate into cells through passive diffusion, 14 different strategies have been explored to obtain intracellular uptake of trehalose. 13 Sonoporation and electroporation can be used for loading trehalose, 13,15,16 but these physical methods could result in excessive cell injuries. 13,17 On the other hand, hydrophobically modified polymers, [17][18][19][20][21] double-layer-charged apatite nanoparticles, 22 and liposomes 23 were also applied for promoting trehalose uptake in cells for cryopreservation or lyophilization.…”

Combining Cooling Enhancement and Trehalose Dehydration to Enable Scalable Volume Cryopreservation of Red Blood Cells with Low Concentration of Glycerol

“…[ 25 ] In particular, after the outbreak of COVID‐19 and the associated containment measures, blood donations decline sharply and cannot meet the demand for routinely used clinical RBCs, highlighting the need for frozen RBCs from another perspective. [ 26 ] Recent advances in the field of RBC cryopreservation have focused on the development of various novel cryoprotectants and intracellular trehalose delivery, including polyampholyte‐based cryoprotective polymers, [ 27,28 ] synthetic nanomaterials with ice recrystallization inhibition properties, [ 29,30 ] polymer mimics of antifreeze proteins, [ 31,32 ] trehalose‐functional glycopeptides, [ 33,34 ] small‐molecule glycosides, [ 35,36 ] natural biocompatible osmoprotectants, [ 37,38 ] membrane‐perturbated trehalose uptake method, [ 39–41 ] sonoporation‐mediated trehalose loading method, [ 42 ] etc. Although these latest scientific findings are stimulating, we would like want to optimize the freezing protocol for RBC cryopreservation in the most straightforward way.…”

“…Furthermore, the functional expression of postwash RBCs after cryopreservation is examined and all results show that this scalable volume cryopreservation method can be extended to current clinical RBC cryopreservation. intracellular trehalose delivery, including polyampholyte-based cryoprotective polymers, [27,28] synthetic nanomaterials with ice recrystallization inhibition properties, [29,30] polymer mimics of antifreeze proteins, [31,32] trehalose-functional glycopeptides, [33,34] small-molecule glycosides, [35,36] natural biocompatible osmoprotectants, [37,38] membrane-perturbated trehalose uptake method, [39][40][41] sonoporation-mediated trehalose loading method, [42] etc. Although these latest scientific findings are stimulating, we would like want to optimize the freezing protocol for RBC cryopreservation in the most straightforward way.…”

Sonoporation of Immune Cells: Heterogeneous Impact on Lymphocytes, Monocytes and Granulocytes

“…Hitherto, no investigation has been dedicated to the analysis of the heterogeneity of leukocyte response to sonoporation. Many MB-US bio-effect investigations on circulatory cells have focused primarily on erythrocytes (i.e., red blood cells) (Miller et al 2000;Chen et al 2003;Janis et al 2021) or cancerous leukemia cells (Miller and Dou 2009;Zhong et al 2011Zhong et al , 2013Duan et al 2021). There have also been scattered studies that have applied MB-US exposure to specific forms of leukocytes such as lymphocytes (Brayman et al 1999;Yong et al 2014;Karki et al 2019), phagocytes (Lemmon et al 2011) and neutrophils (Korosoglou et al 2006).…”