Allostimulatory capacity of conditionally immortalized proximal tubule cell lines for bioartificial kidney application

Abstract

Novel renal replacement therapies, such as a bioartificial kidney (BAK), are needed to improve current hemodialysis treatment of end-stage renal disease (ESRD) patients. As BAK applications may reveal safety concerns, we assessed the alloimmunization and related safety aspects of readily available conditionally immortalized human proximal tubule epithelial cell (ciPTEC) lines to be used in BAK. Two ciPTEC lines, originally derived from urine and kidney tissue, were characterized for the expression and secretion of relevant molecules involved in alloimmunization and inflammatory responses, such as HLA class-I, HLA-DR, CD40, CD80, CD86, as wells as soluble HLA class I and proinflammatory cytokines (IL-6, IL-8 and TNF-α). A lack of direct immunogenic effect of ciPTEC was shown in co-culture experiments with peripheral blood mononuclear cells (PBMC), after appropriate stimulation of ciPTEC. Tight epithelial cell monolayer formation on polyethersulfone flat membranes was confirmed by zonula occludens-1 (ZO-1) expression in the ciPTEC tight junctions, and by restricted inulin-FITC diffusion. Co-culture with (activated) PBMC did not jeopardize the transepithelial barrier function of ciPTEC. In conclusion, the absence of allostimulatory effects and the stability of ciPTEC monolayers show that these unique cells could represent a safe option for BAK engineering application.

A bioartificial kidney device with polarized secretion of immune modulators

“…The concentrations of TNF‐α did not vary in all cases and remained very low (100–300 pg/mL), in agreement with our previous findings (Mihajlovic et al, ; Peters et al, ). If we, however, estimate the absolute amount of TNF‐α in both compartments (based on the volume of each compartment, we observe that the amount of TNF‐α is higher in the extraluminal than in the intraluminal compartment (Figure S3).…”

“…Besides, the high uraemic toxins concentrations in kidney patients are often associated with inflammation, which may as well be detrimental for the BAK (Hsu et al, ). A first assessment of the immunogenicity of the ciPTEC lines using flat membranes with a small surface area (1.12 cm 2 ) showed that ciPTECs have low immunogenicity in vitro, although able to secrete several proinflammatory cytokines that could potentially mediate a non‐specific inflammatory response (Mihajlovic et al, ). In this work, we investigated whether there is polarization of the production of proinflammatory and immune mediators by the ciPTEC cultured in the BAK system.…”

Removal of Mn (II) by Sodium Alginate/Graphene Oxide Composite Double-Network Hydrogel Beads from Aqueous Solutions

“…Since regeneration and reusability are important factors of an adsorbent, plus the high cost of GO based material 58 , the reusability of GAD hydrogel beads in current study need to considered as well. In this experiment, HCl was chosen as desorbent, the adsorbed Mn(II) can be elute with the treatment of 0.1 mol/l HCl.…”

Advances in predictive in vitro models of drug-induced nephrotoxicity

“…The potential utility of IL-6 and IL-8 as in vivo biomarkers of AKI is unclear, but a pilot study that assessed the effects of gentamicin and cadmium chloride on primary proximal tubule cells from two different donors showed greater upregulation of IL-6 and IL-8 than KIM1 levels in response to the compounds 82 . Upregulation of IL-6 and IL-8 expression also occurs in conditionally immortalized proximal tubule cell lines in response to uraemic toxins 93 . Together, these findings suggest that IL-6 and IL-8 are used as readouts of injury in vitro.…”