Abstract: SA-PO689

In Vitro Mast Cell Degranulation Assay to Assess Hemodialysis Sorbent Cartridges

Session Information

Category: Dialysis

  • 604 Home and Frequent Dialysis

Authors

  • Merchant, Stephen, Fresenius Medical Care, Oklahoma City, Oklahoma, United States
  • Costanzo, Mark, Fresenius Medical Care, Oklahoma City, Oklahoma, United States
  • Mullon, Claudy, Fresenius Medical Care, Oklahoma City, Oklahoma, United States
  • Thakuria, Mayuri, Fresenius Medical Care, North America, Waltham, Massachusetts, United States
  • Figueroa, Amparo L, Fresenius, Boston, Massachusetts, United States
  • Kossmann, Robert J., Fresenius Medical Care North America, Waltham, Massachusetts, United States
Background

Hemodialysis (HD) sorbent cartridges aimed at dialysate regeneration contain urease to hydrolyze urea into bicarbonate and ammonium. Urease may come from sources that can cause allergic reactions. Here, we compared in vitro, two sorbents in their propensity to cause allergic reaction.

Methods

In vitro mast cell degranulation method was used to examine dialysate samples from an FDA cleared sorbent cartridge (REDY) and a new cartridge (PAK). Both sorbent cartridges were primed with typical hemodialysis dialysate and recirculated for 30 minutes. Samples for analysis were taken from the recirculated dialysate in the reservoir at 10, 20, and 30 minutes.

Diluted (50 µl) concanavalin A (Con A) standards in complete RPMI plus IL-3 medium (positive controls), culture medium (negative control), or sorbent cartridge effluent test articles were added to 200 µL of culturing medium containing ~0.3 million MC/9 cells, in duplicate. Cells were incubated for 24 hours at 37 degrees C under 5% CO2. Histamine levels in each cell culture supernatant sample were determined using Histamine ELISA Cell Culture Kit. To evaluate the responsiveness of the method, various concentrations of Con A were used as positive controls.

Results

There was no significant difference (p=NS) in the histamine concentrations between both the PAK and REDY sorbent dialysates vs. negative control at 10, 20, and 30 minutes. At Con A concentration of 0.1 – 0.8 µg/ml, positive correlation was observed with histamine release, r=0.95, p<0.001. At con A concentration of 0.1 – 0.8 µg/ml, the mean histamine release was between 17.76 -94.52 ng/ml. On the other hand, the negative control had a mean histamine release of 17.68 ng/ml.

Conclusion

In this proof of concept study, we report the first ever in vitro histamine release testing of sorbent cartridge dialysates incubated in the presence of mast cells. The findings demonstrate that the histamine release from the dialysate generated by the PAK sorbent is not significantly different to that of the negative control culture medium and the dialysate generated by the REDY sorbent.

Funding

  • Commercial Support