Effect of applied encapsulation methods and incubation on proliferation of MSCs post-encapsulation and cryopreservation.

<p>Proliferation of MSCs: (<b>A1)</b> after encapsulation in alginate; (<b>A2</b>) after recovery post-encapsulation; (<b>B1</b>) cryopreserved in alginate; (<b>B2</b>) after recovery post-thawing. “Recovery” – incubation of cells in culture (37°C, 5% CO₂ in a humidified incubator) for 5 days after encapsulation and/or cryopreservation; “Immediate” – immediately after encapsulation; “Incubated” – after 24 h of incubation inside alginate beads in culture; AF – air flow; CF – frozen not encapsulated cells; CN – not encapsulated and not frozen MSCs. The next process parameters were kept constant during encapsulation: both for AF and electro-spraying – spraying distance 10 cm, concentration of gelling solution 100 mM CaCl₂, alginate flow rate 10 ml/h, and alginate concentration 1.6% (w/v); for AF – air flow 150 l/h. In B1–B2 freezing parameters included cooling rate 1 K/min down to −80°C and freezing medium with 10% DMSO and 10% FBS. The data are shown as a mean±SD (n = 3, N = 5). One-way ANOVA: NS – not significant (p = 0.05); *, **, *** - significantly different with p<0.05, p<0.01, p<0.001, respectively.