Review Article

Recent Advances and Future Direction in Lyophilisation and Desiccation of Mesenchymal Stem Cells

Table 2

Different methods to deliver trehalose into cells.

MethodExplanationReference

ElectroporationMurine myeloma cells were loaded with trehalose by electroporation, then freeze-dried, and rehydrated[31]

Genetically engineering the cellsHuman primary fibroblasts were transfected with otsA and otsB genes from E. coli, encoding for trehalose synthase[32]

Genetically engineered poresGenetically engineered mutant of alpha-hemolysin from Staphylococcus aureus was used to create pores in the cellular membrane of 3T3 fibroblasts and human keratinocytes. Resulting nonselective pore is equipped with a metal-actuated switch that is sensitive to extracellular zinc concentrations, thus permitting controlled loading of trehalose[33]

Fluid phase endocytosisHuman MSCs were loaded by trehalose up to 30 mM internal concentration at usual cultivation conditions for 24 h at 37 in the presence of MSC medium with 0–125 mM trehalose[34]

Endogenous cell surface receptorTF-1 cells were permeabilized using an endogenous protein P2X7[35]

Cell penetrating peptides (CPPs)Trehalose was coupled with CPP and incubated with mouse embryonic fibroblast cells at usual cultivation conditions[36]