We have developed a microencapsulation procedure for the entrapment and manipulation of IB3-1 cystic fibrosis cells. The applied method is based on the generation of monodisperse droplets by a vibrational nozzle; for the microbeads hardening, an ionic alginate encapsulation procedure was utilized. Different experimental parameters were analyzed, including frequency and amplitude of vibration, polymer pumping rate and distance between the nozzle and the gelling bath. The produced barium-alginate microbeads posses excellent properties, with respect to morphological parameters as well as narrow size distribution. Results, reproducibly obtained, firmly demonstrate that the microencapsulation procedure do not alter the viability of the encapsulated IB3-1 cells. The encapsulated IB3-1 cells were characterized in term of secretomic profile, analysing the culture medium by Bio-Plex strategy. The analyzed factors include members of the interleukin family, chemokines, growth factors and soluble forms of adhesion molecules. 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-alpha. In this experimental set-up, encapsulated and free IB3-1 cells were treated, for 24 hours, with TNF-alpha, thereafter the culture media from both cell populations were collected. As expected, TNF-alpha induced a sharp increase in the secretion of interleukins, chemokines and growth factors. Of great interest was the evidence that induction of IL-6 and IL-8 occurs also by encapsulated IB3-1 cells. In conclusion, the described encapsulation procedure represents a promising strategy to utilize encapsulation of secreting cells in alginate microbeads, as a very interesting approach for biotechnology applications in tissue engineering and biomedicine.
TNF-alpha induced release of IL-8 by IB3-1 cells encapsulated in alginate microbeads
BORGATTI, Monica;MAZZITELLI, Stefania;BREVEGLIERI, Giulia;GAMBARI, Roberto;NASTRUZZI, Claudio
2009
Abstract
We have developed a microencapsulation procedure for the entrapment and manipulation of IB3-1 cystic fibrosis cells. The applied method is based on the generation of monodisperse droplets by a vibrational nozzle; for the microbeads hardening, an ionic alginate encapsulation procedure was utilized. Different experimental parameters were analyzed, including frequency and amplitude of vibration, polymer pumping rate and distance between the nozzle and the gelling bath. The produced barium-alginate microbeads posses excellent properties, with respect to morphological parameters as well as narrow size distribution. Results, reproducibly obtained, firmly demonstrate that the microencapsulation procedure do not alter the viability of the encapsulated IB3-1 cells. The encapsulated IB3-1 cells were characterized in term of secretomic profile, analysing the culture medium by Bio-Plex strategy. The analyzed factors include members of the interleukin family, chemokines, growth factors and soluble forms of adhesion molecules. 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-alpha. In this experimental set-up, encapsulated and free IB3-1 cells were treated, for 24 hours, with TNF-alpha, thereafter the culture media from both cell populations were collected. As expected, TNF-alpha induced a sharp increase in the secretion of interleukins, chemokines and growth factors. Of great interest was the evidence that induction of IL-6 and IL-8 occurs also by encapsulated IB3-1 cells. In conclusion, the described encapsulation procedure represents a promising strategy to utilize encapsulation of secreting cells in alginate microbeads, as a very interesting approach for biotechnology applications in tissue engineering and biomedicine.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
