Objective
Stem cell-derived islets (SC-islets) represent an important source of cells for the treatment of T1D. As SC-Islets move closer to broad clinical application, scalable and reliable cryopreservation methods will be essential to support manufacturing, storage, and distribution. Optimizing cryopreservation is therefore a critical enabling step in the clinical translation of stem cell-based therapies for diabetes. The proposed investigation endeavors to establish a preservation protocol for SC-Islets. The work starts with characterizing the basic low temperature behavior of the cells in an SC-islet. These studies help define mechanisms of damage during freezing and develop strategies to mitigate that damage. Subsequent studies help determine the composition of cryopreservation solution and method of freezing that results in the highest post thaw recovery and function for the SC-islets. This method can then be tested on SC-islets developed using different manufacturing techniques as well as clinical scale cell numbers.
Background Rationale
Type 1 diabetes mellitus (T1D) is an autoimmune condition in which the body’s immune system targets and destroys pancreatic β-cells. Transplantation of insulin-producing islets has long been recognized as a viable treatment for type 1 diabetes (T1D), demonstrating the potential to achieve insulin independence in select patients. However, widespread application has been limited by the scarcity of donor tissue and the need for chronic immunosuppression. To address these limitations, substantial efforts have focused on generating functional β-cells from renewable sources, such as pluripotent stem cells. Recent studies have shown that induced pluripotent-derived stem cells may restore immune tolerance and help preserve islet β-cell function. As these products move closer to broad clinical application, scalable and reliable cryopreservation methods will be essential to support manufacturing, storage, and distribution. Optimizing cryopreservation is therefore a critical enabling step in the clinical translation of stem cell-based therapies for diabetes.
Diverse biological systems (plants, insects, etc.) survive high salt environments, dehydration, drought, freezing temperatures and other stresses using osmolytes. We have demonstrated our ability to preserve single cells using combinations of osmolytes consisting of sugars, sugar alcohols and amino acids. Tools for understanding events during freezing of cells were traditionally limited to low temperature light microscopy. Low temperature Ramen spectroscopy obtained during freezing provides a rich, molecular-level view of the freezing process including the presence of ice (or not) inside a cell, changes membrane hydration and changes in hydrogen bonding of water molecules in complex solutions. By leveraging this powerful, nondestructive analytical technique, we aim to directly observe and characterize physiochemical changes that occur during cryopreservation. This capability adds greatly to the knowledge available to rationally and scientifically formulate new approaches to cell preservation.
Description of Project
Stem cell-derived islets (SC-islets) represent an important source of cells for the treatment of T1D. As SC-Islets move closer to broad clinical application, scalable and reliable cryopreservation methods will be essential to support manufacturing, storage, and distribution. Optimizing cryopreservation is therefore a critical enabling step in the clinical translation of stem cell-based therapies for diabetes. The proposed investigation endeavors to establish a preservation protocol for SC-Islets. Part of the study involves foundational studies that help elucidate why the cells are damaged during freezing. Subsequent studies help determine the composition of cryopreservation solution and method of freezing that results in the highest post thaw recovery and function for the SC-islets. This work will help us improve preservation of SC-islets.
Anticipated Outcome
The anticipated outcomes of this investigation will be a preservation protocol for stem cell derived islets. Specifically, the protocol will consist of the composition of a cryopreservation solution as well as a method of freezing. The proposed studies include low temperature imaging studies which will help us understand how and why this cell type is damaged during freezing. This knowledge can be used to develop strategies to mitigate that damage and preserve cell viability and function. As a result, these studies will also help us understand more broadly the low temperature behavior of the cells comprising the islet. This knowledge can be used to help us improve the preservation of different cells used therapeutically.
Relevance to T1D
Stem cell derived islets (SC-islets) represent a renewable source of cells for islet transplantation. Recent studies have shown that induced pluripotent-derived stem cells may restore immune tolerance and help preserve islet β-cell function. Furthermore, clinical trials utilizing cGMP-compliant stem cell products have reported beneficial outcomes in T1DM patients, without significant adverse effects. As SC-Islets move closer to broad clinical application, scalable and reliable cryopreservation methods will be essential to support manufacturing, storage, and distribution. Optimizing cryopreservation is therefore a critical enabling step in the clinical translation of stem cell-based therapies for diabetes.