Genomic analysis for hematopoietic stem and progenitors cells (HSPC) generated in vitro according to ex vivo expansion protocols and their comparison with HSPC obtained fresh

Expansion for hematopoietic cells from umbilical cord blood is a strategy for use this cell source in clinic transplants, however, it is important to know about the genomic changes that can occur in expanded cells. In order to detect global expression profiles changes in hematopoietic stem and progenitors cells generated in vitro, we analyzed hematopoietics populations obtained by FACS in fresh from umbilical cord blood. HSC (fHSC) was defined as CD34+ CD38- CD71- CD45RA- Lin- and were cocultured with stromal cell line OP-9 plus FL, SCF, IL3, IL6, TPO, GMCSF and G-CSF by 7 days, after time we repurified HSC population by FACS using same immunophenotype (ivHSC). In other hand, fresh erythroid progenitors cells (fEPC) were identified as CD34+CD38+CD71+CD45RA- Lin- and fresh myeloid progenitors cells (fMPC) were identified as CD34+CD38+CD71-CD45RA+Lin-. In vitro progenitors cells (ivEPC and ivMPC) were obtained by culturing fHSC in Stemspan serum-free media plus SCF, TPO, IL6, FL and IL3 by 10 days, after time cells were repurified by FACS using same immunophenotype for fresh progenitors. In vitro generated cells were compared with their corresponding fresh population cells. Global expressions patterns were compared between generated in vitro populations of Hematopoietic Stem Cells (HSC) and Myeloid Progenitor Cells (MPC), and Erythroid Progenitors Cells (EPC) with HSC, MPC and EPC populations obtained freshly from Umbilical Cord Blood from healthy donors. Each group has at least two or up to four different samples. Populations were purified by FACS and processed in order to obtain those different populations.