Novel Disaccharide Based Cryopreservation Approach for Stable , -80 Degrees C Storage of Stem Cells

ABSTRACT Long term cryopreservation of cells requires the use of cryoprotectant agents CPAs Dimethysulfoxide DMSO is currently the gold standard CPA which has been used for half a century to preserve cells long term While DMSO is considered non toxic to stem cells it does alter cell physiology especially as storage solutions warm More serious is the reported side effects in transplant patients attributed to DMSO toxicity Additional issues with DMSO are requirements for ultralow temperatures which significantly increases the infrastructure needs and costs for storage and shipping We are developing novel injectable CPA alternative to DMSO based on trehalose for banking stem cells as well as other cell types that is also stable at non cryogenic temperatures e g C Trehalose is a non reducing disaccharide that possesses an exceptional ability to stabilize and preserve cells and cellular structures during freezing However unlike DMSO which is freely membrane diffusible trehalose is not naturally taken up by mammalian cells We have overcome this limitation using the ATP activated purinergic receptor P X The work proposed here will refine cryopreservation methods using trehalose and establish optimal freezing and storage parameters If successful this method will significantly reduce the costs and technical hurdles inherent to cryopreservation of many different cell types and tissues NARRATIVE Long term preservation of cells and tissues requires the use of protective chemical substances to prevent injury to cellular structures caused by ice crystal formation during freezing Current agents require ultralow temperatures for long term storage and are associated with toxicities We are developing a storage agent which allows preservation at standard temperatures and is not associated with toxicity to the preserved cells or patients receiving the cells to correct diseases