Hematopoietic stem cells (HSCs) represent a heterogeneous population of primitive blood progenitor cells that fall into several different classes.1,2 Within the human body, they are mainly located in the bone marrow of adults, but are also found in various fetal tissues such as umbilical cord blood, placenta and fetal liver. HSCs are functionally defined by their self-renewal capacity and their multipotency allowing the replenishment of all types of blood cells. The myeloid branch of their descendants is represented by monocytes/macrophages, granulocytes (neutrophils, basophils, eosinophils), erythrocytes, megakaryocytes (platelet producer cells) and dendritic cells. The lymphoid branch comprises of T-lymphocytes, B-lymphocytes and NK-cells.3 Treatments using HSCs (bone marrow transplantation) have been used for over 40 years now and are well-established.4 Indeed, HSCs still hold great potential for further applications in regenerative medicine and are therefore intensively investigated by the scientific community.
It is known that in vivo HSCs are able to expand to very large numbers by virtue of their pronounced self-renewal abilities.5 The in vitro colony forming unit (CFU) assay has been used to study the proliferation and differentiation pattern of hematopoietic progenitors by their ability to form colonies in a semisolid agar medium. However, researchers face difficulties during in vitro expansion of undifferentiated HSCs, with increasing differentiation of the cells observed during the expansion phase in traditional serum-containing culture systems. The establishment of serum-free but undefined formulations provided a somewhat more consistent culture environment, but the results were still not acceptable.
The PromoCell HPC Expansion Medium DXF (C-28021) provides a highly optimized, chemically defined and serum-free culture system for CD34+/CD133+ human hematopoietic stem cells. Optimal support of self-renewal and inhibition of differentiation results in superior expansion of human hematopoietic progenitor cells.
Figure 1.Differentiation of hematopoietic stem cells into various blood and immune cell types.
Figure 2. Expansion of human hematopoietic stem cells. Phase bright morphology and flow cytometry analysis of cord blood derived CD34+ progenitor cells after 12 days of expansion in PromoCell HPC Expansion Medium DXF. More than 98% of the cells are viable after the expansion period as detected by 7-AAD staining.
Figure 3. Characterization of human hematopoietic stem cells. The percentage of hematopoietic stem cells grown in the PromoCell HPC Expansion Medium DXF stain positive for CD34 (>93%), while >95% of the cells do not stain for CD38. The CD34+/CD38- marker profile indicates the maintenance of a primitive hematopoietic phenotype.
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