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  • Encapsulation of RIN-m5F cells within Ba2+ cross-linked alginate beads affects proliferation and insulin secretion.

Encapsulation of RIN-m5F cells within Ba2+ cross-linked alginate beads affects proliferation and insulin secretion.

Journal of microencapsulation (2006-11-23)
Yu-Xin Cui, Kevin M Shakesheff, Gary Adams
ABSTRACT

The viability, proliferation and insulin production of RIN-m5F cells when loaded into alginate beads to form a 3D culture system has been investigated. The mechanism of alginate cross-linking (calcium ions vs barium ions), the addition of poly(L-lysine) (PLL) and poly(L-ornithine (PLO) and presence of different extra-cellular matrix proteins (ECM) influence the RIN-m5F cell behaviour. Cells in calcium alginate beads (CAB) proliferated and produced more insulin per cell than monolayer culture, but the physical properties of the beads were poor and they ruptured within a few days of culture. Barium alginate beads (BABs) provided a stable encapsulation method. Addition of PLL and PLO at concentrations above 0.1% w/v with the culture medium increased cell proliferation. With the addition of ECMs after bead formation there was a further increase in cell proliferation for certain combinations of ECM and PLO. It was concluded that RIN-m5F-loaded Ba-alginate beads (BABs), when incorporated with varying concentrations of poly (L) lysine (PLL), poly (L) ornithine (PLO) in the presence of extra-cellular matrix proteins (ECMs) were superior to both tissue culture and RIN-m5F-loaded Ca-alginate beads (CABs) in terms of physical stability, cell proliferation and insulin production.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly-L-ornithine solution, mol wt 30,000-70,000, 0.01%, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Poly-L-ornithine hydrobromide, mol wt 30,000-70,000
Sigma-Aldrich
Poly-L-ornithine hydrobromide, mol wt 5,000-15,000
Sigma-Aldrich
Poly-L-ornithine hydrobromide, mol wt >100,000