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OMIP-049

Analysis of Human Myelopoiesis and Myeloid Neoplasms

Genyuan Zhu Jason Brayer, Eric Padron, James J. Mulé, Adam W. Mailloux Cytometry PART A, Volume 93, Issue 10, 982-986 (2018)

PURPOSE: Cells undergoing myelopoiesis in both bone marrow and the mature myeloid compartments in the blood represent a broad range of phenotypes in different stages of maturation and activation. Neoplasms that arise from this lineage are often heterogeneous and frequently display unique characteristics and outcomes depending on the originating subset or degree of differentiation 1, 2. This is particularly true for myeloid neoplasms (MN) which rely on multiple, separate flow cytometry panels for differential diagnosis 3, 4. While standard practice, this approach has inherent disadvantages which include the inability to observe co‐expression patterns between markers contained in separate panels and increased time and labor associated with multiple flow cytometry assays. Use of a single, high‐order panel that is able to differentiate multiple hematopoietic lineages, or distinguish different MNs would aid this effort and provide the opportunity to further characterize disease subsets. This panel was designed to detect cell surface markers associated with hematopoiesis with a special emphasis on the progression of myelopoiesis from hematopoietic progenitors. Because the study of hematopoietic disease requires a broad combination of early hematopoietic and mature myeloid markers 5, this panel is also appropriate for the general analysis of myelopoiesis, granulopoiesis, erythropoiesis, and megakaryocytopoiesis, in any human cell source that contains mature myeloid cells, myeloid progenitors, or hematopoietic progenitors (Table 1).

CELL TYPE: Any source containing human myeloid cells

MACHINE: BD LSR2

Marker Target Clone Fluorochrome Purpose
Dead cells Human Live/Dead Fix Aqua Viability
CD138 Human MI15 APC-Cy7 Plasma cell
CD19 Human HIB19 Brilliant Violet 510 exclusion
CD3 Human SK7 Brilliant Violet 510 exclusion
CD45 Human HI30 BUV805 Leukocyte
CD117 Human 104D2 BB515 Progenitor
CD11b Human ICRF44 Brilliant Violet 650 Myeloid Lineage
CD11c Human 3.9 PE-CF594 DC
CD14 Human M5E2 Brilliant Violet 786 Monocyte
CD16 Human 3G8 BUV496 FC receptor
CD33 Human WM53 Brilliant Violet 421 Myeloid Lineage
CD34 Human 581 PE-Cy7 Progenitor
CD36 Human CB38 PerCP-Cy5.5 M2
CD64 Human 10.1 BUV737 Activation, M1
HLA-DR Human G46-6 BUV395 M1
CD15 Human W6D3 Alexa Fluor 700 Myeloid Lineage
CD163 Human GHI/61 Brilliant Violet 605 Macrophage
CD32 Human FUN-2 APC FC receptor
cd41a Human HIP8 PE Megakaryocyte
CD71 Human M-A712 Brilliant Violet 711 Erythroid marker