Laboratery Study Of Ginsenosides, Rg1 And Rb1 On Proliferation And Differentiation Of CD34+ Cell Isolated From Cord Blood In Vitro

Objective: To investigate the effects of ginsenosides, Rgl and Rbl on proliferation and differentiation of CD34+ cell isolated from cord blood ex vivo. Methods:1. CD34+ hematopoietic stem cell was isolated and purified from human cord blood using MACS system. The purity of isolated CD34+ cells were determined by flow cytometry.2. The CD34+ cells were placed in MethoCultTM4435 semisolid medium containing GS, Rgl, Rbl of different concentrations. The numbers of CFU-E, BFU-E and CFU-GM colonies were counted on 7 or 14 day respectively.3. CD34+ cells were plated in StemSpanTMFEM serum-free liquid medium containing 50μg/L TPO and SCF with GS, Rgl, Rbl at different concentrations. Seven days later, numbers of total cell and CD34+ cell were counted respectively. The 7-day-expended cells were placed in MethoCultTM4435 semisolid medium. The numbers of CFU-E, BFU-E and CFU-GM colonies were counted on 7 or 14 day respectively.Results:1. The average purity of CD34+ cells isolated from 10 cord blood samples was about 97%.2. In GS, Rgl, Rbl groups, the numbers of CFU-E, BFU-E, CFU-GM were higher than those in the control group. In general, the effects of Rgl and Rbl were superior to GS. The most effective concentration of GS was 50mg/L, Rgl was 5mg/L, Rbl was 5mg/L. Compared with the best concentrations, in the group of highest concentration of GS(200mg/L), Rgl(50mg/L), Rbl(50mg/L), the numbers of colonies were decreased, but didn't show the inhibitory effect.3. In the liquid culture system, GS, Rgl, Rbl stimulated the proliferation of5CD34+ cell. The most effective concentration of GS was 50mg/L, Rgl was5mg/L, Rbl was 5mg/L. The numbers of total mononuclear cells were 20.4,24.5and 24.1 times higher and CD34+ cell numbers were 4.22,5.19 and 4.12 timeshigher than original numbers. The effects of GS, Rgl, Rbl were significantlyhigher than the control group.4. Colonies formation ability of GS, Rgl, Rbl -treated 7-day expanded cells washigher than that of 7-day expanded control cells. At the concentration of 50mg/LGS, 5mg/L Rgl, 5mg/L Rbl, CFU-E colonies increased 90.1%, 70.8%, 72.8%,BFU-E increased 53.4%, 68.0%, 67.0%, CFU-GM increased 67.0%, 29.8%,39.4%, respectively.Conclusions: GS, Rgl, Rbl could stimulate proliferation and differentiation ofthe hematopoietic stem cell, ginsenosides had analogous and synergetic effectswith the cytokines.