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CD99 as surface anchor for human islet endocrine cell purification.
Geert A Martens1, Veerle De Punt1, Geert Stangé1
1Centre for Beta Cell Therapy in Diabetes, Vrije Universiteit Brussel, Brussels, Belgium.
Journal of Tissue Engineering and Regenerative Medicine
|October 1, 2016
Summary
CD99 is a novel human beta cell surface marker. Its high abundance and trypsin resistance enable efficient magnetic bead-mediated purification of islet endocrine cells for therapeutic applications.
Area of Science:
- Cell biology
- Proteomics
- Immunology
Background:
- Identifying specific cell surface markers is crucial for isolating pure cell populations.
- Beta cells within pancreatic islets play a key role in glucose regulation.
Purpose of the Study:
- To identify and validate a cell surface marker for human beta cell purification.
- To develop a scalable method for enriching functional human islet endocrine cells.
Main Methods:
- Proteomic analysis using LC-MS/MS to identify cell surface proteins.
- Gene and protein expression analysis via microarray, western blotting, flow cytometry, and immunofluorescence.
- Development of a magnetic bead-based cell sorting protocol using anti-CD99 antibodies.
Main Results:
- CD99 was identified as a highly abundant and islet-selective surface protein in human beta cells, but not in rat.
- CD99 expression was confirmed in insulin+/TSQ+ cells and resistant to tryptic digestion.
- Magnetic bead-based purification using anti-CD99 antibodies achieved a fourfold enrichment of functional endocrine cells.
Conclusions:
- CD99 is a reliable human beta cell surface marker.
- CD99 facilitates scalable and efficient purification of islet endocrine cells for potential therapeutic use.
