Abstract: TH-PO396
Inhibition of Erythropoiesis by Soluble Fas in Cell Culture
Session Information
- Cell Signaling and Oxidative Stress
November 02, 2017 | Location: Hall H, Morial Convention Center
Abstract Time: 10:00 AM - 10:00 AM
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.