In Vitro Proliferation Of Human Induced Pluripotent Stem Cells Into Erythroid Colony And Establishment Of A Stable Erythroid Progenitor Cell Line

Objective:To induce human induced pluripotent stem cells (hiPSCs) to differentiate into erythroid progenitor cells through co-culture with trophoblasts in the presence of inducing factors, and establish a stable erythroid progenitor cell line.Method:1. The commercial hiPSCs were induced to differentiate into erythroid colony by the culture system which contain Flt-3, TPO, SCF, EPO, IL-3, and co-culture with CF-1 mouse embryonic fibroblast (MEF) cells.2. The erythroid colonies were passaged to establish a stable erythroid progenitor cell line. The cell morphology was observed by inverted microscope, cell viability determined by MTT assay, cell markers determined by flow cytometry.1. The hiPSC-derived erythroid progenitor cell line grow in colonies, with no significant difference in morphology from the primary cells; while the erythroid progenitor cell line derived from CD34+ cells grow slowly, mostly in suspension and rarely in colonies.2. The Wright staining showed that (72.20±6.24)% of the hiPSC-derived erythroid progenitor cells were positive, while only (35.00±6.02)% of the erythroid progenitor cells derived from CD34+cells were positive. The positive staining rate between the two groups was significantly difference.3. The positive rates of CD71 and CD235a by flow cytometry were (85.09±4.14)% and (5.87±0.37)% respectively for hiPSC-derived erythroid progenitor cells, and (93.52 ±1.59)% and (3.02±0.75)% respectively for primary cells, the differences were significant (P<0.05). The positive rates of CD71 and CD235a were (23.47±1.77)% and (0.58±0.07)% respectively for the erythroid progenitor cells derived from CD34+ cells, and (84.22±3.66)% and (1.02±0.08)% respectively for the primary CD34+cells, the differences were significant (P<0.01). The positive rates of CD71 and CD235a also showed significant differences between the two lines of erythroid progenitor cells (P<0.05).1. The hiPSCs were successfully induced to form erythroid colonies by co-culture with trophoblasts in the presence of inducing factors, and an erythroid progenitor cell line was successfully established.2. Compared with erythroid progenitor cells derived from CD34+ cells, the hiPSC-derived erythroid progenitor cells showed superior inheritance of surface markers and genetic stability.