Affiliations: Hemorheology Center, Department of Biophysics, School of Basic Medical Sciences, Peking University, Beijing 100083, China | Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093-0412, USA
Note: [] Corresponding author: Zongyao Wen, Department of Biophysics, Health Science Center, Peking University, Beijing 100083, P.R. China. Tel.: +86 10 82802419; Fax: +86 10 62015582; E-mail: [email protected].
Abstract: Erythroblasts were obtained from murine spleen. After cultured for 12 hr, the cells were divided into four groups with the use of the following cytokines in culture: EPO, EPO+SCF, EPO+IL-3, and EPO+IL-3+SCF. Cell proliferation assay was done. Apoptosis rates were obtained by using a flow cytometer. Mitochondrial membrane potential (MMP) was assessed in flow cytometry (FCM) by labeling with rhodamine 123. Mitochondrial enzyme activity (MEA) was evaluated with MTT colorimetric assay. The cells were labeled with Fluo-3/Am Ester and Ca2+ concentration was measured. The expression of Bax mRNA and Bcl-2 mRNA was analyzed by RT-PCR. At same time, the expression of Bax and Bcl-2 was analyzed by western blotting. Our results showed that IL-3 and SCF have synergistic effects with EPO on the proliferation, differentiation and apoptosis of erythroid progenitors.
