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Updated: Aug 13, 2026
Human Pancreatic Islet Isolation: Part II: Purification and Culture of Human Islets
Published on: May 26, 2009
Formation of pseudoislets from human pancreatic cultures
C Y Kuo1, H G Herrod, G A Burghen
1University of Tennessee, Pediatric Research, Memphis 38163.
Insights
Researchers developed a method to culture human islet cells from donor pancreata. These cultured cells form pseudoislets that secrete insulin in response to glucose, offering a new model for studying islet function.
Area of Science:
- Endocrinology
- Cell Biology
- Tissue Engineering
Background:
- Human islet cells are crucial for insulin production and diabetes research.
- Current methods for studying islet function in vitro face limitations.
Purpose of the Study:
- To develop a novel in vitro culture technique for human pancreatic islet cells.
- To characterize the behavior and function of cultured human islet cells and derived pseudoislets.
Main Methods:
- Human pancreatic tissue from pediatric cadavers was cultured using collagenase digestion.
- Epithelioid monolayers were formed and stained for insulin.
- Ethylenediaminetetraacetic acid (EDTA) was used to induce pseudoislet formation.
- Insulin secretion was measured in response to varying glucose concentrations.
Main Results:
- Cultured human pancreatic tissue formed insulin-positive epithelioid monolayers within 24-48 hours.
- EDTA treatment yielded spherical pseudoislets with peripheral insulin-positive cells.
- Insulin-positive cells remained detectable and secreted insulin after 3-4 weeks of culture.
- Glucose stimulation induced insulin secretion from these cultured cells.
Conclusions:
- A reproducible in vitro culture technique for human pancreatic islet cells was established.
- The developed pseudoislet model allows for systematic in vitro studies of human islet cell function.
- This method provides a valuable tool for diabetes research and therapeutic development.
Abstract:
We have successfully developed a technique for culturing human islet cells obtained from the cadaveric pancreata of children. Within 24-48 h of in vitro culture, collagenase-digested human pancreatic tissue formed epithelioid monolayers. Scattered within these monolayers were insulin-positive cells, as detected by immunocytochemical and dithizone staining. Treatment of the beta cell-containing epithelioid-cell monolayers with EDTA resulted in the formation of spherical cellular clusters, i.e., pseudoislets. These pseudoislets differed from isolated islets of Langerhans in that they showed a more peripheral distribution of insulin-positive cells. Our studies have demonstrated that insulin-positive cells can be detected in monolayers obtained from human pancreata 3-4 weeks after culture. When exposed to varying concentrations of glucose, these cells secreted insulin. The development of this in vitro technique for culturing human pancreatic islet tissue could provide a model for systematically studying in vitro islet function.
