Biologic and phenotypic analysis of early hematopoietic progenitor cells in umbilical cord blood

Umbilical cord blood (UCB) is an attractive potential alternative to bone marrow (BM) as a source of bematopoietic progenitor cells since the number of progenitors in UCB is similar or even greater than that in normal BM. It was the aim of the present study to analyse the degree of immaturity of UCB progenitor cells. UCB mononuclear (MNC) and/or CD34 ⁺ cells were tested for: a) surface antigen phenotypc; b) expression of cytokines receptor; c) effect of Stem Cell Factor (SCF) on colony growth; d) resistance to mafosfamide and e) replating potential (±SCF). We have found thai 31.3±4.1% and 69.8±3.9% of UCB CD34 ⁺ cells did not express CD38 and CD45RA antigens, respectively, suggesting that UCB contains a high proportion of immature progenitor cells. By means of three color analysis, the receptor for SCF was detected on the majority of the CD34 ⁺ HLA-DR ⁺ subpopulalion; in fact, 76.5±5.1% of CD34 ⁺ HLA-DR ⁺ cells were defined as SCF ^low and 7.6±2.3% as SCF ^high . Colony growth of MNC and CD34 ⁺ cells was enhanced by the addition of SCF to methylcellulose mixture, resulting in a statistically significant increase in CFU-GM (pSO.005) and CFU-GEMM (pSO.005) but not in BFU-E numbers. UCB progenitor cells showed a high resistance to mafosfamide treatment and addition of SCF to the culture medium resulted in a statistically significant increase in mafosfamide concentration required to inhibit 95% of colony growth (pS0.05). Moreover, as shown by single colony transfer assays, the presence of SCF in primary cultures promoted a significantly higher replating potential for both untreated (42±3.3% vs 21±4.6%, pSO.018) and Mafosfamide-treated samples (62±5.6% vs 44±6.1%, pSO.018). In conclusion, UCB is a source of progenitor cells with immature characteristics in terms of surface antigen expression, distribution of SCF receptor, resistance to mafosfamide and replating potential. Therefore, UCB progenitor cells represents an ideal candidate population for experimental programs involving gene transfer and ex vivo stem cell expansion.