Erythropoiesis and Megakaryopoiesis in a Dish

Abstract

Erythrocytes and platelets are the major cellular components of blood. Several hereditary diseases affect the production/stability of red blood cells (RBCs) and platelets (Plts) resulting in anemia or bleeding, respectively. Patients with such disorders may require recurrent transfusions, which bear a risk to develop alloantibodies and ultimately may result in transfusion product refractoriness. Cell culture models enable to unravel disease mechanisms, and to screen for alternative therapeutic products. Besides these applications, the ultimate goal is the large-scale production of blood effector cells for transfusion. Cultured RBCs that lack many of the common blood group antigens and Plts-lacking HLA expression would improve transfusion practice. Large numbers of RBCs and Plts can already be generated using hematopoietic stem cells derived from fetal liver, cord blood, peripheral blood, and bone marrow as starting material for cell culture. The recent advances to generate blood cells from induced pluripotent stem cells provide a donor-independent, immortal primary source for cell culture models. This enables us to study developmental switches during erythropoiesis/megakaryopoiesis and provides potential future therapeutic applications. In this review, we will discuss how erythropoiesis and megakaryopoiesis are mimicked in culture systems and how these models relate to the in vivo process.

Functionalized 3D scaffolds for engineering the hematopoietic niche

“…Nevertheless, 2D systems are easier and more reproducible; moreover, based on the scaffold material, cell collection could be facilitated compared to tridimensional cultures. Hence, they are still preferred for large-scale production and high-throughput screenings ( Bello et al, 2018 ; Varga et al, 2018 ). Moreover, some of the highly promising techniques are represented by microspheroids, organoids, and decellularized tissues (especially natural scaffolds) since they are the most realistic models and facilitate the engraftment of HSCs in vivo ( Yin et al, 2016 ; Allenby et al, 2018 ; Harris et al, 2018 ; Janagama and Hui, 2020 ; Sahu et al, 2021 ).…”

Erythroid Differentiation of Induced Pluripotent Stem Cells Co-cultured with OP9 Cells for Diagnostic Purposes

“…A major limitation of our study is that the cultured erythroid cells remained in the immature RBC state, which may explain the hemoglobin electrophoresis results showing the presence of fetal hemoglobin. Immature RBCs, including fetal RBCs, have been obtained after RBC production from iPSCs in most previ- ous studies [30][31][32][33]. Although blood group antigen expression has been observed in immature RBCs, mature cells are required for a robust reagent RBC development protocol [34].…”

Current status of platelet manufacturing in 3D or bioreactors

“…The strategies for platelet generation consist of HSC amplification, followed by megakaryocyte maturation and differentiation 10 . Since the first report of the in vitro generation of platelets from PB HSCs in 1995, 11 various efforts have been made to optimize protocols using different cell sources.…”

“…The strategies for platelet generation consist of HSC amplification, followed by megakaryocyte maturation and differentiation. 10 Since the first report of the in vitro generation of platelets from PB HSCs in 1995, 11 various efforts have been made to optimize protocols using different cell sources. In combination with thrombopoietin (TPO), which is the major regulator of megakaryopoiesis, other cytokines, including stem cell factor (SCF), feline Mcdonough sarcoma (fms)-like tyrosine kinase 3 (Flt-3), interleukin (IL)-6, IL-9, IL-1β, IL-11 IL-3, Granulocytemacrophage colony-stimulating factor (GM-CSF), erythropoietin (EPO), or other small molecules and proteins, have been added to Cell-groSCGM (CellGenix), StemSpan (Stem Cell Technologies), and IMDMbased media for two or three stages (Table 1).…”