title: Nanoparticle-mediated intracellular delivery enables cryopreservation of human adipose-derived stem cells using trehalose as the sole cryoprotectant.
abstract: In this study, pH responsive genipin-cross-linked Pluronic F127-chitosan nanoparticles (GNPs) was synthesized to encapsulate trehalose for intracellular delivery to cryopreserve primary human adipose-derived stem cells (hADSCs). Trehalose is a disaccharide of glucose used by lower organisms to survive extreme cold in nature and has been used to cryopreserve various biomacromolecules. However, it does not enter mammalian cells because of its highly hydrophilic nature, and has only been used in combination with other cell-penetrating cryoprotectants (such as dimethyl sulfoxide, DMSO) to cryopreserve mammalian cells. Our data show that trehalose can be efficiently encapsulated in our GNPs for intracellular delivery, which enables cryopreservation of primary hADSCs using the nontoxic sugar as the sole cryoprotectant. This capability is important because the conventional approach of cryopreserving mammalian cells using highly toxic (at body temperature) cell-penetrating cryoprotectants requires multistep washing of the cryopreserved cells to remove the toxic cryoprotectant for further use, which is time-consuming and associated with significant cell loss (∼10% during each washing step). By contrast, the trehalose-cryopreserved cells can be used without washing, which should greatly facilitate the wide application of the burgeoning cell-based medicine.
authors: Department of Biomedical Engineering, The Ohio State University , 1080 Carmack Road, Columbus, Ohio 43210, United States.
authors: Wei
authors: W
authors: Rao
authors: Haishui
authors: H
authors: Huang
authors: Hai
authors: H
authors: Wang
authors: Shuting
authors: S
authors: Zhao
authors: Jenna
authors: J
authors: Dumbleton
authors: Gang
authors: G
authors: Zhao
authors: Xiaoming
authors: X
authors: He
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D039823:Iridoids / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry* / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D020442:Poloxamer / Q000737:chemistry; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014199:Trehalose / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity
chemical_list: D003451:Cryoprotective Agents; D018960:Hyaluronan Receptors; D039823:Iridoids; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014157:Transcription Factors; D020442:Poloxamer; C007834:genipin; D014199:Trehalose
keywords: cryopreservation; genipin; nanoparticle; stem cells; trehalose