Characterization of induced pluripotent stem cell‐derived megakaryocyte lysates for potential regenerative applications

Abstract

Recently, platelet‐derived growth factors present in lysates became an extreme interest in the field of regenerative medicine such as in wound healing and as substitutes to foetal bovine serum in xeno‐free cell culture systems. However, the generation of such platelet lysates completely depends on the availability of platelet donors. In this study, the possibility to use in vitro‐generated megakaryocytes derived from induced pluripotent stem cells (iPSCs) as a cell source for typical platelet growth factors was investigated. Therefore, the presence and levels of those factors were characterized in in vitro‐produced megakaryocytes. In comparison with platelets, in vitro‐generated megakaryocytes showed a multifold increased content in transcript and protein levels of typical platelet growth factors including platelet‐derived growth factors (PDGFs), transforming growth factor (TGF)‐1β, vascular endothelial cell factor (VEGF)‐A, epidermal growth factor (EGF), insulin‐like growth factor (IGF)‐1 and tissue factor (TF). Hence, iPSC‐derived megakaryocytes may serve as an efficient cell source for a donor‐independent generation of growth factor‐rich lysates with a broad application potential in innovative cell culture systems and regenerative therapies.

Human‐induced pluripotent stem cell‐derived blood products: state of the art and future directions

“…CD41a is also expressed on hematopoietic progenitor cells and should therefore always be combined with CD42 expression to show megakaryoid commitment. Although high percentages of CD41a are obtained by multiple groups (50-99%), only the inducible cMYC, BMI1 and BCL-XL imMKCLs model achieved almost complete CD42b positivity (98-99.8%; others 30-60%) [11,27,54,55,[57][58][59]62]. Alongside this surface marker combination, polyploidization is another hallmark of MK maturation.…”

“…However, in contrast, other systems, such as micro carrier culture or 2D differentiation have shown a superior MK yield of between~30 and 6900 MK/iPSC [11,27,62]. However, in contrast, other systems, such as micro carrier culture or 2D differentiation have shown a superior MK yield of between~30 and 6900 MK/iPSC [11,27,62].…”

“…Although high percentages of CD41a are obtained by multiple groups (50-99%), only the inducible cMYC, BMI1 and BCL-XL imMKCLs model achieved almost complete CD42b positivity (98-99.8%; others 30-60%) [11,27,54,55,[57][58][59]62]. Different cell-surface markers correspond to MK maturation, among which are CD41a and CD42.…”

“…The embryoid body-based method is a commonly applied initiation step in iPSC-MK differentiation. However, in contrast, other systems, such as micro carrier culture or 2D differentiation have shown a superior MK yield of between~30 and 6900 MK/iPSC [11,27,62]. These systems improved the iPSC-MK output substantially, even compared to CD34 + cultures yielding 10-20 MK/CD34 + cells [27].…”

“…iPSC-MKs exhibit a broad range and degree of polyploidization. Within CD41a + cells, 38%-55% are > 4N, whereas, based on CD42b, this percentage has a broader spread of between 6% and 51% [55,[57][58][59][60]62]. This makes it difficult to assess whether this variation in polyploidization is caused by biological (ontogeny) or technical (differentiation method) factors.…”