Abstract: TH-PO396

Inhibition of Erythropoiesis by Soluble Fas in Cell Culture

Session Information

Category: Cell Biology

  • 203 Growth Factors, Chemokines, Autacoids

Author

  • Goes, Miguel A, Wake Forest University, Winston Salem, North Carolina, United States
Background

Serum soluble Fas (sFas) levels are associated with anemia in CKD patients. Cord blood cells (CBc) can generate hematopoietic stem cells (CD34+). Objective: To investigate whether sFas interferes with erythropoiesis in cell culture

Methods

We studied CD34+-cell culture after sorting from CBc. Analyzed on flow cytometry for glycophorin A, CD 34+, CD 38+ and CD 171+. The CD34+ cells were incubated for 14 days in methylcellulose medium containing growth factors added with sFas or without sFas. They were divided in 18 wells from 6 plates. We divided in 2 groups. Each group consisted in 9 wells with different concentrations of sFas (H group- [2, 4 and 8 ng/mL]; L group-[0, 0.5, and 1 ng/mL]). We performed counting of colony-forming unit (CFU) numbers in all wells for red series (B/CFU-E), white series (CFU-GM) and mixed series (CFU-GEMM) under inverted light microscopy. We performed correlations and comparisons.

Results

We found that CD34+ was the major marker after sorting. We observed a negative correlation between sFas concentration and B/CFU-E (r=-0.74; p<0.001) when analyzed all 18 wells together. We observed lower amount of B/CFU-E in plate with sFas-8ng/ml than in the plates with sFas-1ng/ml, 0.5 ng/ml and non-sFas, 3.7+1.1, 4.3+2.0, 5+1.7, respectively; p=0.02). There was a negative correlation between B/CFU-E and CFU-GM (r=-0.69; p=0.03) in the H-CD34+ group. We observed lower number of B/CFU-E in H-CD34+ group than in the L-CD34+ group (1.5+0.9, 4.3+1.6; p=0.008).

Conclusion

We observed that higher concentrations of sFas interfere with erythropoiesis in CD34+ cell culture.