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Proliferation of sorted human and rat beta cells
- Source :
- Diabetologia, Vol. 51, No 1 (2008) pp. 91-100
- Publication Year :
- 2008
-
Abstract
- AIMS/HYPOTHESIS: The aim of the study was to determine whether purified beta cells can replicate in vitro and whether this is enhanced by extracellular matrix (ECM) and growth factors. METHODS: Human beta cells were purified by FACS by virtue of their high zinc content using Newport Green, and excluding ductal and dead cells. Rat beta cells were sorted by autofluorescence or using the same method developed for human cells. Cells were plated on poly-L-lysine or ECMs from rat or human bladder carcinoma cells or bovine corneal ECM and incubated in the presence of BrdU with or without growth factors. RESULTS: The newly developed method for sorting human beta cells yields a population containing 91.4 +/- 2.8% insulin-positive cells with a low level of spontaneous apoptosis and a robust secretory response to glucose. Beta cells from 8-week-old rats proliferated in culture and this was increased by ECM. Among growth factors, only human growth hormone (hGH) and the glucagon-like peptide-1 analogue liraglutide enhanced proliferation of rat beta cells, with a significant increase on both poly-L-lysine and ECM. By contrast, sorted adult human beta cells from 16 donors aged 48.9 +/- 14.3 years (range 16-64 years) failed to replicate demonstrably in vitro regardless of the substratum or growth factors used. CONCLUSIONS/INTERPRETATION: These findings indicate that, in our conditions, the fully differentiated human adult insulin-producing beta cell was unable to proliferate in vitro. This has important implications for any attempt to expand cells from pancreases of donors of this age group. By contrast, the rat beta cells used here were able to divide in vitro, and this was enhanced by ECM, hGH and liraglutide.
- Subjects :
- Adult
medicine.medical_specialty
Insulin-Secreting Cells/cytology/physiology
Adolescent
Endocrinology, Diabetes and Metabolism
Cellular differentiation
medicine.medical_treatment
Population
Cell Culture Techniques
Islets of Langerhans Transplantation
Cell Culture Techniques/methods
Cell Separation
Biology
Extracellular matrix
Diabetes Mellitus/therapy
Islets of Langerhans Transplantation/methods
Species Specificity
Internal medicine
Insulin-Secreting Cells
Internal Medicine
medicine
Diabetes Mellitus
Animals
Humans
ddc:576.5
education
Beta (finance)
education.field_of_study
ddc:617
Growth factor
Pancreatic islets
Cell Differentiation
Middle Aged
Flow Cytometry
Flow Cytometry/methods
In vitro
Cell biology
Extracellular Matrix
Rats
Endocrinology
medicine.anatomical_structure
Beta cell
Subjects
Details
- Language :
- English
- ISSN :
- 0012186X
- Database :
- OpenAIRE
- Journal :
- Diabetologia, Vol. 51, No 1 (2008) pp. 91-100
- Accession number :
- edsair.doi.dedup.....bfedecb02576011b595f2937fab94ded