Authors:

Valentina Andrade Pérez

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About Paper:

Hematopoietic stem cells (HSCs) are widely recognized as promising candidates for therapeutic interventions in hematological and immunological disorders. Current scientific investigations are primarily dedicated to the creation of robust platforms and reliable techniques for effective differentiation of human pluripotent stem cells (hPSCs) into HSCs, to overcome the limitations associated with allogenic HSCs. Here, we show that HSCs can be efficiently obtained from a 3D cell culture of hPSCs by modulating Wnt and TGFβ signaling pathways via small molecules. To initiate the differentiation process, we used the hanging drop method to induce the formation of embryoid bodies (EBs) from hPSCs. Subsequently, we treated these EBs with vascular endothelial growth factor (VEGF), CHIR99021 (a Wnt activator) and SCF and FLT3L, along with SB431542 (a TGFβ signaling inhibitor) to induce HSC differentiation. The resulting HSCs were treated with GlutaMax, granulocyte stimulating factor (G-SCF) and AM-580 to further differentiate into neutrophils. Immunostaining analysis was performed to monitor the differentiation progress of EBs into HSCs and then neutrophils, which showed the presence of HSC makers within the EBs after 13 days of treatment. These findings suggest that Wnt agonists and TGFβ inhibitors can promote the differentiation of 3D cell culture of hPSCs into HSCs. Future studies should focus on optimizing and characterizing this differentiation protocol to enhance the efficiency and functionality of the derived HSCs.

Source:

Purdue University / 2023

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Co-authors:

Valentina Andrade Pérez