Transplantation of stem cell-derived β cells is a promising treatment for type-1 diabetes, increasing the supply of insulin-producing cells beyond that of cadaveric islet transplantation. Transplant success is limited by cell death after transplantation, with insufficient oxygen and nutrient accessibility strongly contributing to apoptosis and de-differentiation. Herein, we investigate cotransplantation of endothelial cells and fibroblasts with stem cell-derived β cells to enhance survival and function posttransplantation. A microporous poly (lactide coglycolide) scaffold was used for culture and transplantation of stem cell-derived β cells. Coculture of the stem cell-derived β cells with endothelial cells and fibroblasts generated vascular networks during in vitro culture, which persisted through transplantation and enhanced in vivo vascularization. 7-days of in vitro culture supported enhanced survival of transplanted cells, though function in terms of insulin secretion and reduction of hyperglycemia was compromised. However, 3-days of preculture led to both improved survival and function of the stem cell-derived β cells, with transplant recipients demonstrating reduced fasting blood glucose levels. These studies demonstrate the potential and some constraints on the application of vascularization strategies to enhance function of stem cell-derived β cells with transplantation to extrahepatic sites.
Impact Statement
Cell death after transplantation due to hypoxia is a significant barrier to cell replacement therapy for type 1 diabetes. While pancreatic islets are highly vascularized, stem cell-derived β cells lack any vasculature and thus are highly susceptible to dysregulation or cell death upon transplantation. In order to enhance engraftment and thus improve transplanted cell function, stem cell-derived β cells can be cocultured with vasculogenic cells to form microvascular networks in vitro that persist during transplantation and anastomose with host vasculature.
KrentzNAJ, SheaLD, HuisingMO, et al.Restoring normal islet mass and function in type 1 diabetes through regenerative medicine and tissue engineering. Lancet Diabetes Endocrinol, 2021; 9(10):708–724; doi: 10.1016/S2213-8587(21)00170-4
2.
KalraS, MukherjeeJ, VenkataramanS, et al.Hypoglycemia: The neglected complication. Indian J Endocrinol Metab, 2013; 17(5):819–834; doi: 10.4103/2230-8210.117219
3.
PambiancoG, CostacouT, EllisD, et al.The 30-Year Natural History of Type 1 Diabetes Complications. Diabetes. Diabetes, 2006; 55(5):1463–1469; doi: 10.2337/db05-1423
4.
ShapiroAMJ, LakeyJRT, RyanEA, et al.Islet Transplantation in Seven Patients with Type 1 Diabetes Mellitus Using a Glucocorticoid-Free Immunosuppressive Regimen. N Engl J Med, 2000; 343(4):230–238; doi: 10.1056/NEJM200007273430401
5.
EmamaulleeJA, ShapiroAMJ. Factors Influencing the Loss of β-Cell Mass in Islet Transplantation. Cell Transplant, 2007; 16(1):1–8; doi: 10.3727/000000007783464461
6.
OgawaY, NomaY, DavalliAM, et al.Loss of glucose-induced insulin secretion and 6LUT2 expression in transplanted 5-cells. 1995.
7.
DemineS, SchiavoAA, Marín-CañasS, et al.Pro-inflammatory cytokines induce cell death, inflammatory responses, and endoplasmic reticulum stress in human iPSC-derived beta cells. Stem Cell Res Ther, 2020; 11(1):7; doi: 10.1186/s13287-019-1523-3
8.
AamodtKI, PowersAC. Signals in the pancreatic islet microenvironment influence β‐cell proliferation. Diabetes Obes Metab, 2017; 19(Suppl 1):124–136; doi: 10.1111/dom.13031
9.
BurganovaG, BridgesC, ThornP, et al.The Role of Vascular Cells in Pancreatic Beta-Cell Function. Front Endocrinol (Lausanne), 2021; 12:667170; doi: 10.3389/fendo.2021.667170
10.
JanssonL, CarlssonP-O. Graft vascular function after transplantation of pancreatic islets. Diabetologia, 2002; 45(6):749–763; doi: 10.1007/s00125-002-0827-4
11.
KaleA, RogersNM. No time to die—how islets meet their demise in transplantation. Cells, 2023; 12(5):796; doi: 10.3390/cells12050796
12.
NaziruddinB, IwahashiS, KanakMA, et al.Evidence for instant blood-mediated inflammatory reaction in clinical autologous islet transplantation. Am J Transplant, 2014; 14(2):428–437; doi: 10.1111/ajt.12558
13.
DesaiNM, GossJA, DengS, et al.Elevated portal vein drug levels of sirolimus and tacrolimus in islet transplant recipients: Local immunosuppression or islet toxicity? 1: Transplantation, 2003; 76(11):1623–1625; doi: 10.1097/01.TP.0000081043.23751.81
14.
KawaharaT, KinT, KashkoushS, et al.Portal vein thrombosis is a potentially preventable complication in clinical islet transplantation. Am J Transplant, 2011; 11(12):2700–2707; doi: 10.1111/j.1600-6143.2011.03717.x
15.
DelauneV, BerneyT, LacotteS, et al.Intraportal islet transplantation: The impact of the liver microenvironment. Transpl Int, 2017; 30(3):227–238; doi: 10.1111/tri.12919
16.
ErikssonO, EichT, SundinA, et al.Positron emission tomography in clinical islet transplantation. Am J Transplant, 2009; 9(12):2816–2824; doi: 10.1111/j.1600-6143.2009.02844.x
17.
MeierJJ, Hong-McAteeI, GalassoR, et al.Intrahepatic transplanted islets in humans secrete insulin in a coordinate pulsatile manner directly into the liver. Diabetes, 2006; 55(8):2324–2332; doi: 10.2337/db06-0069
18.
GiblyRF, ZhangX, LoweWL, et al.Porous scaffolds support extrahepatic human islet transplantation, engraftment, and function in mice. Cell Transplant, 2013; 22(5):811–819; doi: 10.3727/096368912X636966
19.
HlavatyKA, GiblyRF, ZhangX, et al.Enhancing human islet transplantation by localized release of trophic factors from PLG scaffolds. Am J Transplant, 2014; 14(7):1523–1532; doi: 10.1111/ajt.12742
20.
KasputisT, CloughD, NotoF, et al.Microporous polymer scaffolds for the transplantation of embryonic stem cell derived pancreatic progenitors to a clinically translatable site for the treatment of type I diabetes. ACS Biomater Sci Eng, 2018; 4(5):1770–1778; doi: 10.1021/acsbiomaterials.7b00912
21.
YoungbloodRL, SampsonJP, LebiodaKR, et al.Microporous scaffolds support assembly and differentiation of pancreatic progenitors into β-cell clusters. Acta Biomater, 2019; 96:111–122; doi: 10.1016/j.actbio.2019.06.032
22.
LiF, CrumleyK, BealerE, et al.Fas ligand-modified scaffolds protect stem cell derived β-cells by modulating immune cell numbers and polarization. ACS Appl Mater Interfaces, 2023; 15(44):50549–50559; doi: 10.1021/acsami.2c12939
23.
BealerE, CrumleyK, CloughD, et al.Extrahepatic transplantation of 3D cultured stem cell-derived islet organoids on microporous scaffolds. Biomater Sci, 2023; 11(10):3645–3655; doi: 10.1039/D3BM00217A
24.
BarkaiU, WeirGC, ColtonCK, et al.Enhanced oxygen supply improves islet viability in a new bioartificial pancreas. Cell Transplant, 2013; 22(8):1463–1476; doi: 10.3727/096368912X657341
25.
ColtonCK. Oxygen supply to encapsulated therapeutic cells. Adv Drug Deliv Rev, 2014; 67–68:93–110; doi: 10.1016/j.addr.2014.02.007
26.
LiangJ-P, AccollaRP, SoundirarajanM, et al.Engineering a macroporous oxygen-generating scaffold for enhancing islet cell transplantation within an extrahepatic site. Acta Biomater, 2021; 130:268–280; doi: 10.1016/j.actbio.2021.05.028
27.
ForsterNA, PeningtonAJ, HardikarA, et al.A prevascularized tissue engineering chamber supports growth and function of islets and progenitor cells in diabetic mice. Islets, 2011; 3(5):271–283; doi: 10.4161/isl.3.5.15942
28.
ChendkeGS, KharbikarBN, AsheS, et al.Replenishable prevascularized cell encapsulation devices increase graft survival and function in the subcutaneous space. Bioeng Transl Med, 2023; 8(4):e10520; doi: 10.1002/btm2.10520
29.
InoguchiK, AnazawaT, FujimotoN, et al.Impact of prevascularization on immunological environment and early engraftment in subcutaneous islet transplantation. Transplantation, 2024; 108(5):1115–1126; doi: 10.1097/TP.0000000000004909
30.
BoT, PascucciE, CapuaniS, et al.3D bioprinted mesenchymal stem cell laden scaffold enhances subcutaneous vascularization for delivery of cell therapy. Biomed Microdevices, 2024; 26(3):29; doi: 10.1007/s10544-024-00713-2
31.
Ben-ShaulS, LandauS, MerdlerU, et al.Mature vessel networks in engineered tissue promote graft–host anastomosis and prevent graft thrombosis. Proc Natl Acad Sci U S A, 2019; 116(8):2955–2960; doi: 10.1073/pnas.1814238116
32.
VlahosAE, KinneySM, KingstonBR, et al.Endothelialized collagen based pseudo-islets enables tuneable subcutaneous diabetes therapy. Biomaterials, 2020; 232:119710; doi: 10.1016/j.biomaterials.2019.119710
33.
ChenX, AlediaAS, GhajarCM, et al.Prevascularization of a fibrin-based tissue construct accelerates the formation of functional anastomosis with host vasculature. Tissue Eng Part A, 2009; 15(6):1363–1371; doi: 10.1089/ten.tea.2008.0314
34.
AghazadehY, PoonF, SarangiF, et al.Microvessels support engraftment and functionality of human islets and hESC-derived pancreatic progenitors in diabetes models. Cell Stem Cell, 2021; 28(11):1936–1949.e8; doi: 10.1016/j.stem.2021.08.001
35.
NalbachL, RomaLP, SchmittBM, et al.Improvement of islet transplantation by the fusion of islet cells with functional blood vessels. EMBO Mol Med, 2021; 13(1):e12616; doi: 10.15252/emmm.202012616
36.
ChenS, WangW, ShenL, et al.A 3D-printed microdevice encapsulates vascularized islets composed of iPSC-derived β-like cells and microvascular fragments for type 1 diabetes treatment. Biomaterials, 2025; 315:122947; doi: 10.1016/j.biomaterials.2024.122947
37.
HogrebeNJ, AugsornworawatP, MaxwellKG, et al.Targeting the cytoskeleton to direct pancreatic differentiation of human pluripotent stem cells. Nat Biotechnol, 2020; 38(4):460–470; doi: 10.1038/s41587-020-0430-6
38.
UrbanczykM, ZbindenA, Schenke-LaylandK. Organ-specific endothelial cell heterogenicity and its impact on regenerative medicine and biomedical engineering applications. Adv Drug Deliv Rev, 2022; 186:114323; doi: 10.1016/j.addr.2022.114323
39.
StaelsW, De GroefS, HeremansY, et al.Accessory cells for β‐cell transplantation. Diabetes Obes Metab, 2016; 18(2):115–124; doi: 10.1111/dom.12556
40.
StaelsW, HeremansY, HeimbergH, et al.VEGF-A and blood vessels: A beta cell perspective. Diabetologia, 2019; 62(11):1961–1968; doi: 10.1007/s00125-019-4969-z
41.
FriendNE, RiojaAY, KongYP, et al.Injectable pre-cultured tissue modules catalyze the formation of extensive functional microvasculature in vivo. Sci Rep, 2020; 10(1):15562; doi: 10.1038/s41598-020-72576-5
42.
CarterJ, KarmiolS, NagyM, et al.Pretransplant islet culture: A comparison of four serum-free media using a murine model of islet transplantation. Transplant Proc, 2005; 37(8):3446–3449; doi: 10.1016/j.transproceed.2005.09.073
43.
ShindoY, KalivarathanJ, SaravananPB, et al.Assessment of culture/preservation conditions of human islets for transplantation. Cell Transplant, 2022; 31:9636897221086966; doi: 10.1177/09636897221086966
44.
BöyükG, YiğitAA. Effects of media supplements on viability and functionality in long-term cultured islet cells. J Appl Biol Sci, 2022; doi: 10.5281/ZENODO.6590591
45.
LingZ, HannaertJC, PipeleersD. Effect of nutrients, hormones and serum on survival of rat islet beta cells in culture. n.d.
46.
JaramilloM, BanerjeeI. Endothelial cell co-culture mediates maturation of human embryonic stem cell to pancreatic insulin producing cells in a directed differentiation approach. J Vis Exp, 2012(61):3759; doi: 10.3791/3759
47.
SeoH, SonJ, ParkJ-K. Controlled 3D co-culture of beta cells and endothelial cells in a micropatterned collagen sheet for reproducible construction of an improved pancreatic pseudo-tissue. APL Bioeng, 2020; 4(4):46103; doi: 10.1063/5.0023873
48.
Kaufman-FrancisK, KofflerJ, WeinbergN, et al.Engineered vascular beds provide key signals to pancreatic hormone-producing cells. Gerecht S. ed. PLoS One, 2012; 7(7):e40741; doi: 10.1371/journal.pone.0040741
49.
JohanssonU, RasmussonI, NiclouSP, et al.Formation of composite endothelial cell–mesenchymal stem cell islets. Diabetes, 2008; 57(9):2393–2401; doi: 10.2337/db07-0981
50.
WeizmanA, MichaelI, Wiesel-MotiukN, et al.The effect of endothelial cells on hESC-derived pancreatic progenitors in a 3D environment. Biomater Sci, 2014; 2(11):1706–1714; doi: 10.1039/C4BM00304G
51.
PagetMB, MurrayHE, BaileyCJ, et al.Rotational co-culture of clonal β-cells with endothelial cells: Effect of PPAR-γ agonism in vitro on insulin and VEGF secretion. Diabetes Obes Metab, 2011; 13(7):662–668; doi: 10.1111/j.1463-1326.2011.01392.x
52.
DavalliAM, ScagliaL, ZangenDH, et al.Vulnerability of Islets in the Immediate Posttransplantation Period. Diabetes, 1996; 19.
53.
LiG, Craig-SchapiroR, RedmondD, et al.Vascularization of human islets by adaptable endothelium for durable and functional subcutaneous engraftment. Sci Adv, 2025; 11(5):eadq5302; doi: 10.1126/sciadv.adq5302
54.
MooreSJ, Gala-LopezBL, PepperAR, et al.Bioengineered stem cells as an alternative for islet cell transplantation. World J Transplant, 2015; 5(1):1–10; doi: 10.5500/wjt.v5.i1.1
55.
EinsteinSA, SteynLV, WeegmanBP, et al.Hypoxia within subcutaneously implanted macroencapsulation devices limits the viability and functionality of densely loaded islets. Front Transplant, 2023; 2:1257029; doi: 10.3389/frtra.2023.1257029
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