Abstract
We have developed a microencapsulation procedure for the entrapment and manipulation of IB3-1 cystic fibrosis cells. The applied method is based on generation of monodisperse droplets by a vibrational nozzle. Different experimental parameters were analyzed, including frequency and amplitude of vibration, polymer pumping rate and distance between the nozzle and the gelling bath. We have found that the microencapsulation procedure does not alter the viability of the encapsulated IB3-1 cells. The encapsulated IB3-1 cells were characterized in term of secretomic profile, analyzing the culture medium by Bio-Plex strategy. The experiments demonstrated that most of the analyzed proteins, were secreted both by the free and encapsulated cells, even if in a different extent. In order to determine the biotechnological applications of this procedure, we determined whether encapsulated IB3-1 cells could be induced to pro-inflammatory responses, after treatment with TNF-α. In this experimental set-up, encapsulated and free IB3-1 cells were treated with TNF-α, thereafter the culture media from both cell populations were collected. As expected, TNF-α induced a sharp increase in the secretion of interleukins, chemokines and growth factors. Of great interest was the evidence that induction of interleukin-6 and interleukin-8 occurs also by encapsulated IB3-1 cells.
Highlights
Cystic fibrosis (CF) is an autosomal recessive disorder caused by mutations of the CF transmembrane conductance regulator (CFTR) gene, which encodes a transmembrane protein present on a variety of cell types and organelles [1]
The excess of mucus is largely caused by the influx of neutrophils, attracted to the site by the increased expression of chemokines such as interleukin-6 (IL-6) [5] and interleukin-8 (IL-8) [5, 6], by bacterial products and inflammatory cytokines
Summarizing, the current paper describes the following: (a) the encapsulation of IB3-1 cells in alginate microbeads following a microencapsulation procedure developed in our laboratory that is based on the generation of monodisperse droplets by an air-driven droplet generator for cell encapsulation; (b) the characterization of alginate microbeads produced, with different experimental parameters, including atomizing air, polymer pumping rate, and distance between the nozzle and the gelling bath; (c) the determination of viability of the encapsulated IB3-1 cells, and (d) the characterization of the encapsulated IB3-1 cells, in terms of secretomic profile, analyzing the culture medium by BioPlex strategy [18,19,20]
Summary
Cystic fibrosis (CF) is an autosomal recessive disorder caused by mutations of the CF transmembrane conductance regulator (CFTR) gene, which encodes a transmembrane protein present on a variety of cell types and organelles [1]. While CF is classically characterized by the presence of pancreatic insufficiency and recurrent lung infections in infants, a wide clinical spectrum has been identified in adults [2, 3]. The excess of mucus is largely caused by the influx of neutrophils, attracted to the site by the increased expression of chemokines such as interleukin-6 (IL-6) [5] and interleukin-8 (IL-8) [5, 6], by bacterial products and inflammatory cytokines. IL-8 is induced transcriptionally by a wide variety of stimuli including tumor necrosis factor-alpha (TNF-α), hyperosmotic shock, and bacterial products such as lipopolysaccharides [9]
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