Fig. 1

Illustrates the guidance of human hematopoietic stem cell (HSC) differentiation toward specialized immune lineages with human bone marrow-derived mesenchymal stem cells (BM-MSCs) and CD3⁺ T cells. A Human CD34⁺HSCs (1 × 10⁵) were isolated from three donors and cultured alone or co-cultured with allogenic MSCs (2 × 10⁴) and activated stem cell donor-matched T cells (1 × 10.⁵) (HSCs/MSCs/T) for 14 days ex vivo. After that, cells were harvested, and their total RNA was extracted to examine the expression of specific genes using quantitative real-time PCR. RNA from HSCs alone, MSCs alone, and T cell alone was evenly mixed in equal amounts as the control group sample, HSCs + MSCs + T. The expression levels of representative genes, including S100A8 (Granulocyte lineage), IL-7R (Lymphoid lineage), GATA-1 (Erythroid lineage), and TMF-1 (Megakaryocyte lineage), are depicted as folds of the control level. Suspensive HSC cells alone and the combination of (HSCs + MSCs + T) from three various HSC donors were harvested at 14 days, and the CD11b⁺ MPO⁺ population was identified using flow cytometry (B). The statistical analysis of ex vivo differentiation from three different donors is summarized in C. Data were expressed as mean ± S.E.M., and significant differences between groups were indicated (*p < 0.05, **p < 0.01; two-sided unpaired t test, ns, not significant)