Bone marrow cell suspensions were stained with a cocktail of PE-conjugated antibodies to Lin positive markers (CD3, CD11b, CD45R, Gr-1, F4/80, CD11c, Ter-119) and with antibodies to CD117 (c-Kit, PE-Cy7), SCA- 1 (APC), CD34 (FITC), and CD16/CD32 (PcP-Cy5). Myeloid progenitors (common myeloid progenitors [CMP], granulocyte-macrophage progenitors [GMP], megakaryocyte-erythroid progenitors [MEP]) were discriminated based on the expression of CD34 and CD16/CD32 within the gate of Lin–CD117+ cells. Common lymphoid progenitors were identified according to the expression of interleukin-7Ra+ (IL-7Ra+) within the Lin− gate. Reagents are shown in supplemental Table 2. Cells were detected using the BD FACSCanto II, BD LSRFortessa or BD Accuri C6 and analyzed with BD FACSDiva and FlowJo (9.3.2) software.
To evaluate LT-HSC (Long-Term Hematopoietic Stem Cells), ST-HSC (Short-Term Hematopoietic Stem Cells), and MMPs (Multipotent progenitors), we used the following Abs: CD34, CD16/32. CD41, Sca-1, CD150, CD48, CD105, and c-Kit all obtained from BD. The classification of the different MPPs was performed according to Pietras et al.31 (link)
To evaluate LT-HSC (Long-Term Hematopoietic Stem Cells), ST-HSC (Short-Term Hematopoietic Stem Cells), and MMPs (Multipotent progenitors), we used the following Abs: CD34, CD16/32. CD41, Sca-1, CD150, CD48, CD105, and c-Kit all obtained from BD. The classification of the different MPPs was performed according to Pietras et al.31 (link)
