TY - JOUR
T1 - Pancreatic islet cryopreservation by vitrification achieves high viability, function, recovery and clinical scalability for transplantation
AU - Zhan, Li
AU - Rao, Joseph Sushil
AU - Sethia, Nikhil
AU - Slama, Michael Q.
AU - Han, Zonghu
AU - Tobolt, Diane
AU - Etheridge, Michael
AU - Peterson, Quinn P.
AU - Dutcher, Cari S.
AU - Bischof, John C.
AU - Finger, Erik B.
N1 - Funding Information:
J.S.R. was supported by Schulze Diabetes Institute and Division of Transplantation at the Department of Surgery, University of Minnesota. The authors thank F. Zhou and W. Zhang from Characterization Facility of the University of Minnesota for help with TEM and C. Forster and A. Lewis of the University of Minnesota Clinical and Translational Science Institute for technical assistance with histology experiments. The authors acknowledge Schulze Diabetes Institute, Department of Surgery for use of their confocal microscope and for providing porcine islets. This work is supported by grants from Regenerative Medicine Minnesota (E.B.F. and J.C.B.), the National Science Foundation (EEC 1941543, J.C.B. and E.B.F.) and the National Institutes of Health (R01DK131209, J.C.B. and E.B.F.; R01DK117425, J.C.B. and E.B.F.; and R01HL135046, J.C.B. and E.B.F.). L.Z. acknowledges the Doctoral Dissertation Fellowship from the University of Minnesota, and J.C.B. acknowledges support from the Kuhrmeyer Chair in Mechanical Engineering and the Bakken Chair in the Institute for Engineering in Medicine from the University of Minnesota. P.P.Q. would like to thank the J.W Kieckhefer Foundation, the Stephen and Barbara Slaggie Family, and the Khalifa Bin Zayed Al Nahyan Foundation for supporting this work.
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/4
Y1 - 2022/4
N2 - Pancreatic islet transplantation can cure diabetes but requires accessible, high-quality islets in sufficient quantities. Cryopreservation could solve islet supply chain challenges by enabling quality-controlled banking and pooling of donor islets. Unfortunately, cryopreservation has not succeeded in this objective, as it must simultaneously provide high recovery, viability, function and scalability. Here, we achieve this goal in mouse, porcine, human and human stem cell (SC)-derived beta cell (SC-beta) islets by comprehensive optimization of cryoprotectant agent (CPA) composition, CPA loading and unloading conditions and methods for vitrification and rewarming (VR). Post-VR islet viability, relative to control, was 90.5% for mouse, 92.1% for SC-beta, 87.2% for porcine and 87.4% for human islets, and it remained unchanged for at least 9 months of cryogenic storage. VR islets had normal macroscopic, microscopic, and ultrastructural morphology. Mitochondrial membrane potential and adenosine triphosphate (ATP) levels were slightly reduced, but all other measures of cellular respiration, including oxygen consumption rate (OCR) to produce ATP, were unchanged. VR islets had normal glucose-stimulated insulin secretion (GSIS) function in vitro and in vivo. Porcine and SC-beta islets made insulin in xenotransplant models, and mouse islets tested in a marginal mass syngeneic transplant model cured diabetes in 92% of recipients within 24–48 h after transplant. Excellent glycemic control was seen for 150 days. Finally, our approach processed 2,500 islets with >95% islets recovery at >89% post-thaw viability and can readily be scaled up for higher throughput. These results suggest that cryopreservation can now be used to supply needed islets for improved transplantation outcomes that cure diabetes.
AB - Pancreatic islet transplantation can cure diabetes but requires accessible, high-quality islets in sufficient quantities. Cryopreservation could solve islet supply chain challenges by enabling quality-controlled banking and pooling of donor islets. Unfortunately, cryopreservation has not succeeded in this objective, as it must simultaneously provide high recovery, viability, function and scalability. Here, we achieve this goal in mouse, porcine, human and human stem cell (SC)-derived beta cell (SC-beta) islets by comprehensive optimization of cryoprotectant agent (CPA) composition, CPA loading and unloading conditions and methods for vitrification and rewarming (VR). Post-VR islet viability, relative to control, was 90.5% for mouse, 92.1% for SC-beta, 87.2% for porcine and 87.4% for human islets, and it remained unchanged for at least 9 months of cryogenic storage. VR islets had normal macroscopic, microscopic, and ultrastructural morphology. Mitochondrial membrane potential and adenosine triphosphate (ATP) levels were slightly reduced, but all other measures of cellular respiration, including oxygen consumption rate (OCR) to produce ATP, were unchanged. VR islets had normal glucose-stimulated insulin secretion (GSIS) function in vitro and in vivo. Porcine and SC-beta islets made insulin in xenotransplant models, and mouse islets tested in a marginal mass syngeneic transplant model cured diabetes in 92% of recipients within 24–48 h after transplant. Excellent glycemic control was seen for 150 days. Finally, our approach processed 2,500 islets with >95% islets recovery at >89% post-thaw viability and can readily be scaled up for higher throughput. These results suggest that cryopreservation can now be used to supply needed islets for improved transplantation outcomes that cure diabetes.
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U2 - 10.1038/s41591-022-01718-1
DO - 10.1038/s41591-022-01718-1
M3 - Article
C2 - 35288694
AN - SCOPUS:85126266062
VL - 28
SP - 798
EP - 808
JO - Nature Medicine
JF - Nature Medicine
SN - 1078-8956
IS - 4
ER -