Speaker
Description
Introduction
Diabetes is a chronic, globally prevalent disease characterized by impaired pancreatic islet cell function. Existing treatments, such as pancreatic islet transplants or exogenous insulin, do not fully restore physiological pancreatic function. This has prompted the search for alternative therapies. Implementing 3D bioprinting of functional pancreatic organoids that mimic native islet architecture and function can improve disease modeling (e.g., diabetes, pancreatic cancer), toxicity assessment, and drug testing without animal use. These models also hold promise for replacement therapy, reconstructing damaged tissues rather than only treating symptoms. In the future, commercial cells may be replaced by patient-derived ones for personalized models and therapy testing. This represents a major opportunity for bioengineering advancement.
The study aimed to evaluate the functionality of 3D printed pancreatic organoids intended to mimic islets of Langerhans..
Methods
Bioprinted 3D pancreatic organoids were made using electromagnetic inkjet printing technology. The bioink used for printing was based on hydrogels derived from methacrylated gelatin (GELMA) and hyaluronic acid (HAMA) and a cross-linking agent (LAP). A cell suspension from four cell lines was added to the bioink, these were the following cell lines: alpha cells (αTC-1) and beta cells (βTC-tet) of pancreatic islets, endothelial cells (HUVEC) and fibroblast cells (L929) in different concentrations depending on the proposed variant. Six proposed variants of tested organoids:
V1:αTC-1:βTC-tet (ratio 1:2),
V2:αTC-1:βTC-tet (ratio 1:3),
V3:αTC-1:βTC-tet:HUVEC(ratio 1:2:3),
V4:αTC-1 βTC-tet:HUVEC (ratio 1:2:1),
V5:αTC-1:βTC-tet:HUVEC: L929 (ratio 1:2:1:2),
V6:αTC-1:βTC-tet:HUVEC : L929 (ratio 2:4:1:1).
The organoids were cultured in standard conditions for 28 days. Biological analyses determining the functionality of the models consisted of: assessment of viability (FDA/Pi method), microscopic evaluation, H&E staining and immunohistochemistry (glucogon, GSIS (insulin), CD31, vimentin) and immunohistochemical tests.
Discussion
The obtained results demonstrate that 3D bioprinted organoids have significant potential in tissue engineering, especially for supporting diabetes patients by mimicking native islet functions. They create functional islets with insulin-producing β cells, reflecting natural structures. This is a major breakthrough, as previous animal models are limited and classical cell cultures fail to replicate complex tissue architecture. Organoids allow for more realistic testing and will enable personalized medicine by transforming pluripotent stem cells (iPSCs) into α, β, endothelial, and fibroblast cells of a patient. This supports precise disease modeling and therapy testing—a major advance in precision medicine.
Results
In all variants (V1-V6) of bioprinted 3D pancreatic organoids their viability was confirmed for 28 days. In the case of V5 and V6, the earliest spheroid formation was observed (7 days of the experiment). Immunohistochemistry of preparations from individual variants showed the activity of α, β cells, endothelium and fibroblasts. Histological imaging showed that in the tested models spheroids are spontaneously formed from alpha and beta cells.
Additionally, the GSIS test showed insulin secretion in all tested variants, the highest concentration on day 28 was shown by V6 (170 ng/ml for high glucose). For variants that did not contain endothelium and fibroblasts (V1 and V2), half the secretory value of β cells was observed. The organoids are functional and show the activity of pancreatic islets.
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