Ex vivo culture of hematopoietic stem and progenitor cells with platelet lysate: Investigating proliferation and erythroid-megakaryocytic lineage effects

Author:

Farnaz Pirsavabi, Zahra Najafi , Mohammad Ebrahimi Siahboomi , Bewar Othman Hasan, Shaban Alizadeh , Seyyed Hadi Mousavi , Mostafa Saberian , Zahra Kashani Khatib, Saeed Mohammadi , Naser Ahmadbeigi, Azadeh Omidkhoda

:Abstract

Hematopoietic stem and progenitor cells (HSPCs) are valuable for therapies and research due to their self-renewal and differentiation abilities. The ex vivo culture of these cells necessitates conditions that maintain their unique properties and provide a niche-like environment. Human platelet lysate (HPL), a growth factor-rich substance used in stem cell studies, influences various biological pathways. This study investigates HPL’s effects on HSPCs’ proliferation and erythroid-megakaryocytic differentiation, addressing the need to improve HSPC culture conditions for clinical and research applications. HPL was prepared, and isolated peripheral blood-derived HSPCs were cultured for 7 days in control and 10 % HPL-supplemented groups. Cell counts, viability, and population doubling time (PDT) were measured. Erythroid and megakaryocytic differentiation was assessed using qPCR and flow cytometry. Colony-forming cell (CFC) assays confirmed multilineage differentiation capacity. HPL significantly increased HSPCs proliferation, resulting in a 1.96-fold increase by day 7 and a reduced PDT. CFC assays confirmed that the cells maintained their ability to differentiate into various blood cell types. Flow cytometry showed a decrease in the CD71 marker without affecting the overall red blood cell population. Gene analysis revealed increased β-globin production while other red blood cell-related genes remained stable. Markers and genes associated with megakaryocyte development showed no significant changes. HPL as a culture medium supplement markedly enhances the proliferation of peripheral blood-derived hematopoietic stem cells during cell culture without inducing unwanted bias into the erythroid and megakaryocytic lineages.