Table 1 Multicellular beta-cell assemblies and their applications. CCL22: C-C motif chemokine ligand 22. CHPOA: Acrylate modified, cholesterol bearing pullulan. ECM: Extracellular matrix. GLP-1: Glucagon-like peptide-1. GPCR: G-protein coupled receptor. hiPSC: human induced pluripotent stem cell. IL: interleukin. MBP-FGF2: Maltose-binding protein-basic fibroblast growth factor 2. MSC: Mesenchymal stem cell. PEG: Poly(ethylene glycol). PEI: Poly(ethyleneimine). PDMS: Polydimethylsiloxane. PNIPAAm: Poly(N-isopropylacrylamide). PMMA: Poly(methyl methacrylate). STZ: Streptozotocin. T1D: Type 1 diabetes.

Spheroid

• 100 mm² non-adherent plastic dish

• ECM gel (laminin, collagen type IV, heparin sulfate proteoglycan, entactin)

MIN6

Spheroids as models to control unlimited growth after transplantation

[235]

Spheroid

PDMS-based concave molds coated with bovine serum albumin

Dissociated rat islet and hepatocyte

Relationship between islets and hepatocytes for cell-based liver or T1D therapy

[236]

Spheroid

Hanging drop

Dissociated rat islet

Assembly of nonviral PEI–mediated IL-10 cytokine gene-delivered cells into spheroids

[237]

Spheroid

3D clinostat

(microgravity)

MIN6

Optimization of spheroid properties for transplantation potential

[238]

Pseudoislet

Spheroid

96-well non-adhesive prime surface plate

α-TC1 clone 6, NIT-1 and TGP52

Effects of seeding density and cell type ratio on cell distribution, hormone mRNA level and functionality

[239]

Spheroid

6-well plate with PNIPAAm-coated PDMS-based microwells

NIT-1

Therapeutic potential in STZ-induced diabetic mouse

[240]

Spheroid

PDMS-based microwell coated with PNIPAAm

MIN6 and MAEC

MIN6 and NIH3T3

Encouragement of insulin secretion induction with companion cells

[241]

Spheroid

100-mm tissue culture dish

INS-1E expressing insulin-GLase

Example case to analyze protein secretion via video-rate bioluminescence imaging

[242]

Spheroid

Layer-by-layer coating with fibronectin and gelatin followed by low cell attachment 96-well U-bottom plate

MIN6

Cell coating for structural stability and functionality of spheroids in T1D treatment

[243]

Spheroid

Membrane-bottomed

PDMS concave microwell array

Dissociated rat islet

Development of a novel system to overcome oxygen supply limitation for functional and viable spheroid formation

[244]

Spheroid

Microfluidic chip containing hemispheric cell culture chambers and with methyl cellulose and collagen type I containing medium feed

Human pancreatic 1.1B4

Development of a platform with perfusion flow network that enables beta-cell spheroid assembly and measurement of beta-cell function under dynamic conditions

[245]

Spheroid

Ultra-low attachment 100 mm dish

MIN6-K20

Model along with monolayer culture in the study of incretin responsiveness state of beta-cells

[246]

Spheroid

Geometry-controlled hanging drop

βTC6

Effects of droplet spreading on the spheroid formation and function

[247]

Spheroid

Microfluidic chip containing PDMS-based concave microwells

Dissociated rat islet

Comparison of static culture with dynamic culture, drug testing (tolbutamide, GLP-1, rapamycin)

[115]

Spheroid

PDMS microwell coated with PNIPAAm or collagen I

MIN6 and NIH3T3

Alteration of cell localization by control of migration

[248]

Spheroid

PDMS-chip and PMMA-chip

MIN6,

MIN6-m9

Study of the role of oxygen supply and antioxidant mediated minimization of reactive oxygen species during bioartificial pancreas preparation

[249]

Spheroid

Hanging drop

MIN6 and rat bone marrow MSC

Development of heterospheroid model for functional and survival boost combined with PEG hydrogel as immunoisolation barrier

[250]

Spheroid

Concave microwell coated with bovine serum albumin

Human liver cells transdifferentiated into insulin producing cell

Spheroid size control, improvement of actin–myosin-associated cytoskeleton changes mediated differentiation and function in vitro and STZ-induced diabetic mouse

[251]

Spheroid

• 35 mm non-adherent petri dish

• Hanging drop

• Agarose 3D microwell technique

• Spherical plate 5D

Dissociated rat islet

Comparison of different spheroid assembly techniques for obtaining optimum islet mimics

[252]

Spheroid

Honeycomb polygons of PDMS

hiPSC

Cellular and functional characterization, effects of dimension, potential to establish pancreas-on-chips

[253]

Spheroid

PDMS honeycomb sheet coated with pluronic acid

hiPSC

Transcriptomics and metabolomics profile comparison of spheroids in culture of 3D static vs. microfluidic chip

[254]

Spheroid

Stirred-tank bioreactor

INS-1

Effects of technique and stirrer type in the assembly, viability, functionality and yield

[255]

Spheroid

Ultra-low binding round-bottom plate as part of differentiation process

MBP-FGF2 immobilized surface as part of differentiation process

Human omentum-derived stem cell

In vitro differentiation in the spheroid formation platform, characterization, and analysis of therapeutic potential in STZ-induced diabetic mouse

[256]

Spheroid

Organoid

Amikagel containing 96-well plate during aggregation following final differentiation

Human embryonic stem cell derived pancreatic progenitor cell and HUVEC

Development and validation of a hydrogel-based system for progenitor cell differentiation towards pancreatic lineage and organoid assembly

[257]

Spheroid

Organoid

Agarose-patterned

microwell

Dissociated islet and human amniotic epithelial cell

Enhancement of engraftment, viability, and graft function in STZ-induced diabetic mouse

[258]

Organoid

Microfluidic chip with microwell configurations as part of differentiation process

hiPSC lines

iPSCs as a source of insulin-producing beta-cells for diverse uses

[259]

Organoid

Agarose-coated 96-well plate

Dissociated mouse islet and mouse microvascular fragment

In vitro characterization and development to address challenge of insufficient vascularization after islet transplantation

[260]

Organoid (or pseudoislet)

Ultra-low attachment 96-well microplate

Dissociated mouse islet

Assessment of spheroid potential for genetic manipulation with adeno-associated virus serotype 8

[261]

SC-islet (organoid)

Ultra-low attachment cell culture plate on orbital shaker

Human H1

Model beta-cell system for the development of the reverse phase liquid chromatography-tandem mass spectrometry for insulin detection

[262]

Pseudoislet

Tissue culture flask coated with gelatin

MIN6

Potential of E-cadherin on the regulation of pseudoislet growth

[263]

Pseudoislet

90 mm bacterial (uncoated) petri dish

MIN6

Development of an immunoisolation barrier for cell-based therapy in T1D

[264]

Pseudoislet

Uncoated 90 mm petri

dishes

MIN6 and mouse MSC

Testing layer-by-layer nanoencapsulation system on the spheroids as models for immunoisolation

[265]

Pseudoislet

6-well tissue culture plate

βTC3 and MS1

Effect of dimension and endothelial cells on function, ECM synthesis and viability

[266]

Pseudoislet

Ultra-low attachment, 6-well, flat-bottomed plate

Human 1.1B4

Evaluation of pseudoislets as alternatives to natural islets for transplantation to STZ-induced diabetic mouse

[267]

Pseudoislet

Agarose gel-based microwell

Dissociated rat islet

Characterization to investigate usefulness of pseudoislets for transplantation into STZ-induced diabetic mouse

[268]

Pseudoislet

2D clinostat

(microgravity) followed by culture in 6 cm petri dish

EndoC-βH3

Description of response to hypoxia

[269]

Pseudoislet

Hanging drop

MIN6

Optimization of alternative beta-cell model and development of CHPOA nanogel coating for immunoprotection

[270]

Pseudoislet

96-well spheroid microwell plate

Dissociated human islet

Characterization including cellular heterogeneity and assessment of spheroid potential for genetic manipulation with lentivirus

[271]

Pseudoislet

Centrifugal forced aggregation in 24-well or 6-well AggreWell 400 plate

Dissociated human islet

Potential to be an alternative for native islets and superiority upon transplantation in STZ-induced diabetic mouse

[272]

Pseudoislet

96-well ultra-low adherent culture plate

Alpha-, beta- or delta-cells from dissociated human islet

Addressing beta-cell source limitation in the replacement therapy by reprogrammed alpha- and delta-cells

[273]

Pseudoislet

Hanging drop

Dissociated human islet

Study of effects of glucose, adrenaline and palmitate on the regulation of insulin and glucagon secretion

[274]

Pseudoislet

Hanging drop

(GravityPLUS)

Dissociated mouse islet

Assessment of spheroid potential by adenovirus-delivered V1b receptor overexpression in vitro and STZ-induced diabetic mouse

[275]

Pseudoislet

6-well tissue culture plate

βTC3 and MS1

MIN6 and MS1

Effect of dimension, endothelial cells and secretagogues on function and gene expression

[276]

Pseudoislet

Ultra-low attachment microplate

Hanging drop (GravityPLUS)

Dissociated human islet

Spheroid cell content, viability, functionality (dynamic perfusion and microfluidic system) analysis to develop representative model in GPCR signaling analysis

[277]

Pseudoislet

96-well plate with a non-adherent round bottom

EndoC-βH3

Monitoring and analysis of endocrine activity in the dynamic system of a microfluidic chip that allows self-trapping of pseudoislets

[278]

Pseudoislet

Suspension flask with stirring

INS832/3

Characterization of insulinoma pseudoislets

[279]

Pseudoislet

Agarose-coated microwell array

αTC1 clone 6, INS-1E and HUVEC

Analysis of cell distribution within spheroids

[280]

Pseudoislet

PDMS microfluidic system with microtrap chambers

α-TC1 clone 6 and INS-1E

Islet-on-a-chip microfluidic 3D model for mimicking in vivo for spheroid assembly and for drug test platform for palmitic acid hydroxy stearic acid

[281]

Pseudoislet

Low attachment microwell plate coated with pluronic acid

αTC1 clone 6 and INS-1E

Model in the discovery of the compounds effective on the reduction of oxidative stress with viability and functionality assays

[282]

Pseudoislet

96-well plates with V-bottom conical wells pre-treated with anti-adherence rinsing solution

Mouse and human FAC-sorted beta-cell and HUVEC

Analysis of impact of beta-cell CD63 expression in transplantation

[283]

Pseudoislet

AggreWell 400 6-well plate followed by centrifugation and shaking, transfer to an ultralow adsorption 6-well plate

MIN6 and HUVEC

Elucidate underlying mechanism of insulin secretion augmentation in the presence of HUVECs, investigation of beta-cell markers and, analysis of therapeutic potential in STZ-induced diabetic mouse

[284]

Pseudoislet

Agarose mold

βTC3 and RHMVEC

Spheroid characterization and endothelial sprouting in fibrin hydrogel to facilitate islet revascularization and function

[285]

Pseudoislet

Magnetic levitation

EndoC-βH3 and HUVEC

Effects of beta-cell and endothelial cell distribution and cellular interactions on beta-cell activity

[286]

Pseudoislet

Spheroid

Hanging drop

MIN6 and hepatic/pancreatic stellate cell

Investigation of heterospheroid culture parameters on beta-cells, genetic engineering and testing stellate cells to express CCL22 to attract Tregs in vivo

[287]

Pseudoislet

Spheroid

Non-adherent culture dishes followed by rotary cell culture reactor with high aspect ratio vessel (HARV)

MIN6 and PANC-1, PaTu-8988t, FA-6

Beta-cell and pancreatic cancer cell interaction

[288]

Pseudoislet

Spheroid

Organoid

AggreWell 400 24-well plate

Dissociated rat islet, HUVEC and hAEC

In vitro characterization, xenogeneic transplantation to facilitate vascularization and engraftment

[289]

Aggregate

Ultra-low attachment 60 mm diameter dish

Dissociated rat islet

Comparison of natural islet and reaggregated islet response against oxidative stress and function encapsulated within alginate microcapsule and beta-cell replacement in STZ-induced diabetic mouse

[290]

Aggregate

Culture medium dependent differentiation and aggregation

Human dental pulp stem cell

As a source for autologous stem cell therapy in diabetes

[291]

Aggregate

Culture medium dependent differentiation and aggregation

Human adipose tissue derived stem cell

As a source for autologous stem cell therapy in STZ-induced diabetic mouse

[292]

Aggregate

PEG microwell followed by 2 h on orbital shaker and culture under static cultures

MIN6

Defining optimum size with functional beta-cell mass

[293]

Aggregate

Ultra-low attachment 24-well plate on 3D rotator

MIN6

Mechanism of superior functional maintenance in 3D configuration

[294]

Aggregate

Non-adherent agarose microwell

Dissociated human islets and commercial murine cell lines

Optimization of spheroid properties to minimize mass transport limitation and, validation in vivo

[295]

Aggregate

6-well plate on a gyratory shaker

αTC-6 and βTC-1

Mimic heterogeneity and cellular communication

[296]

Aggregate

Hanging drop

Mouse sertoli cell and mouse dissociated islet cell

Development of cell-based immunosuppressive therapy in beta-cell replacement in STZ-induced diabetic mouse

[297]

Aggregate

96-well culture plate with V-bottomed wells

INS-1 and gelatin hydrogel microspheres

Effects of microspheres on beta-cell viability and functionality

[298]

Aggregate

Non-adherent agarose microwell chip

MIN6-B1 and HUVEC

Study of beta-cell function within membrane-based encapsulation devices and effect of endothelial cells

[299]

Aggregate

Gelatin–PLGA porous microwell scaffolds

RIN-5F

Effects of dimensionality and microwell size on aggregation and function of beta cells

[300]

Cluster

12-well microplate

Dissociated rat islet cells transfected to express exendin-4

Investigation of a novel strategy for diabetes treatment with aggregates of genetically engineered islet cells, PEG-lipid coating and anti-CD154 mAb and tacrolimus supplementation

[301]

Cluster

Culture medium dependent differentiation and aggregation

Human adipose tissue derived stem cell

Characterization of a beta-cell source in stem cell therapy in diabetes

[302]

Cluster

Hanging drop

Dissociated rat islet

Potential to be an alternative for native islets and superiority upon transplantation with matrigel in STZ-induced diabetic mouse

[303]

Cluster

Culture medium dependent differentiation and aggregation in 6-well plates on an orbital shaker

Human pluripotent stem cell

As a source for stem cell therapy with dynamic response and in diabetes in vivo

[304]

Cluster

Microwell bag

hiPSC line (1231A3 and Ff-I14s04) derived insulin secreting cell

Advancement of scalability, sterility and operability

[305]

Cluster

AggreWell 400/800 microwells as a part of differentiation protocol

Human

pluripotent stem cell

Improvement of stem-cell derived beta-cell establishment protocol for diverse applications such as small molecule screening

[306]

Cluster

Spheroid

Hanging drop

Dissociated rat islet and human bone marrow MSC

Capacity of MSCs to benefit in islet transplantation outcome by angiogenesis and anti-apoptosis

[307]

Cluster

Spheroid

Hanging drop

Dissociated rat islet and curcumin-loaded PLGA microspheres

Ability of beta-cell clusters to overcome hypoxia-induced death of transplantation through curcumin

[308]

Vascularized islet

Low cell-adhesion

96-well plate with U-bottomed conical wells

Islet, HUVEC and human MSC

Development of vascularized islet organoid transplants to improve therapeutic efficacy

[309]

Islet-like structure

During culture medium dependent differentiation (incubation with protamine chloride)

Human liver stem-like cell

Endocrine differentiation into islet-like structures and beta-cell replacement potential in STZ-induced diabetic mouse

[310]

SC-islet

As a part of culture medium dependent differentiation (6-well AggreWell 400 plates followed by ultra-low attachment plates)

Human H1

Detailed characterization from cytoarchitecture to insulin secretory machinery

[311]

Stem-cell derived islet

AggreWell 400 plates and vertical-wheel bioreactors as a part of differentiation protocol

hiPSC

Generation and economical optimization of islet-like cluster with bioreactor included system and, characterization as well as analysis of effective islet transplantation mass

[312]

Islet microtissue

96-well hanging drop system

Dissociated human islet

As a model to determine the protective capacity of liraglutide on GSIS, cytokine secretion and T cell infiltration in islet microtissue-PBMC co-cultures

[313]