IN VITRO RESPONSE OF UT-7 CELL LINE TO ERYTHROPOIETIN-RESISTANT AND ERYTHROPOIETIN-RESPONSIVE SERUM FROM HEMODIALYSIS PATIENTS IN ADDINGTON & KING EDWARD VIII HOSPITALS, KWAZULU NATAL- SOUTH AFRICA

8 Feb 2025 12 a.m. 12 a.m.
WCN25-AB-2867, Poster Board= SAT-314

Introduction:

Erythropoietin (EPO) therapy is widely used to correct anemia in hemodialysis (HD) patients. However, a subset of these patients, termed EPO-resistant, requires significantly higher doses of EPO to achieve adequate hemoglobin levels. This study aims to investigate the role of serum in EPO resistance among HD patients, categorized as EPO-resistant, EPO-responsive, and healthy controls.

Methods:

The study evaluated the impact of serum from HD patients with varying EPO responsiveness on the proliferation of UT-7 cells, which are EPO-dependent. UT-7 cells (50,000 cells per well) were cultured in triplicate in a 96-well plate with either no serum (control) or serum from EPO-resistant patients (Hb < 10 g/dL despite 20,000 IU weekly EPO), EPO-responsive patients (Hb > 10 g/dL with 12,000 IU weekly EPO), or healthy controls. The plates were incubated in a CO2 environment for 24 hours. Cell proliferation was assessed by measuring ATP levels using a GloRunner Microplate Luminometer (Turner Biosystem, Sunnyvale, CA, USA) with a Cell Titer Glo assay. Luminescence, recorded as relative light units (RLU), was converted to ATP concentrations using a standard curve (y = 0.0004x + 0.383). Statistical analysis was conducted using SPSS version 21 (IBM).

Results:

The bioassay quantified cell proliferation and viability in the presence of EPO. UT-7 cells cultured without serum exhibited significantly higher ATP production (p < 0.0097) compared to cells cultured with serum from any of the three groups (Figure 1). No statistically significant difference was observed in the ATP production of UT-7 cells cultured with serum from EPO-resistant patients, EPO-responsive patients, or healthy controls.

Conclusions:

The addition of serum to UT-7 cell cultures was associated with reduced ATP production, indicating that serum may contain inhibitory substances that limit cell proliferation. Although serum from EPO-resistant patients showed the greatest reduction in ATP production, this difference was not statistically significant. The observed reduction in ATP production suggests that factors such as cytokines or hepcidin in the serum may contribute to diminished cell proliferation. Further research is needed to explore the lack of significant differences between EPO-responsive and EPO-resistant sera and healthy control sera, as this study's findings are based on a limited sample size.

 

Response of serum of hemodialysis patients and healthy controls on UT-7 cell proliferation.

     

I have no potential conflict of interest to disclose.

I did not use generative AI and AI-assisted technologies in the writing process.