Robust generation of erythroid and multilineage hematopoietic progenitors from human iPSCs using a scalable monolayer culture system

Ruiz, Juan Pablo Pizarro; Chen, Guibin; Haro‐Mora, Juan J.; Keyvanfar, Keyvan; Liu, Chengyu; Zou, Jizhong; Beers, Jeanette; Bloomer, Hanan; Qanash, Husam; Uchida, Naoya; Tisdale, John F.; Boehm, Manfred; Larochelle, André

doi:10.1016/j.scr.2019.101600

Cited by 25 publications

(38 citation statements)

References 61 publications

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“…Cells have even been generated through processes that recapitulate in vitro the EHT that takes place in the AGM as a way to ensure acquisition of stemness to produce pluripotent HSCs with the capacity to colonize hematopoietic sites. However, these cells still lack a robust capacity to engraft in a host (Wang et al, 2005;Lengerke et al, 2009;Doulatov et al, 2013;Riddell et al, 2014;Ruiz et al, 2019;Zhou et al, 2019). The first experiment that succeeds to produce engraftable HSCs from iPS cells achieved this by driving differentiation in vivo through teratoma formation, but even in this case, the efficiency was very low with less than 1-2% of engraftment and relied on complex procedures and cocktails of cytokines to achieve this (Amabile et al, 2013;Suzuki et al, 2013;Philipp et al, 2018).…”

Section: In Vitro Hematopoietic Transdifferentiation From Ips/es Cellmentioning

confidence: 99%

Multifaceted Actions of GFI1 and GFI1B in Hematopoietic Stem Cell Self-Renewal and Lineage Commitment

Beauchemin

Möröy

2020

Front. Genet.

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Section: In Vitro Hematopoietic Transdifferentiation From Ips/es Cellmentioning

confidence: 99%

Multifaceted Actions of GFI1 and GFI1B in Hematopoietic Stem Cell Self-Renewal and Lineage Commitment

Beauchemin

Möröy

2020

Front. Genet.

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“…More recently, established iPSCs from human fibroblast cells represent a powerful tool for the investigation of early hematopoiesis [ 59 , 76 , 77 ]. One of the promising strategies for the use of iPSC is their capacity to differentiate into RBCs and to eliminate the allogeneic blood shortages [ 78 , 79 ].…”

Section: Introductionmentioning

confidence: 99%

Differentiation of human induced pluripotent stem cells into erythroid cells

et al. 2020

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During the last years, several strategies have been made to obtain mature erythrocytes or red blood cells (RBC) from the bone marrow or umbilical cord blood (UCB). However, UCB-derived hematopoietic stem cells (HSC) are a limited source and in vitro large-scale expansion of RBC from HSC remains problematic. One promising alternative can be human pluripotent stem cells (PSCs) that provide an unlimited source of cells. Human PSCs, including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), are self-renewing progenitors that can be differentiated to lineages of ectoderm, mesoderm, and endoderm. Several previous studies have revealed that human ESCs can differentiate into functional oxygen-carrying erythrocytes; however, the ex vivo expansion of human ESC-derived RBC is subjected to ethical concerns. Human iPSCs can be a suitable therapeutic choice for the in vitro/ex vivo manufacture of RBCs. Reprogramming of human somatic cells through the ectopic expression of the transcription factors (OCT4, SOX2, KLF4, c-MYC, LIN28, and NANOG) has provided a new avenue for disease modeling and regenerative medicine. Various techniques have been developed to generate enucleated RBCs from human iPSCs. The in vitro production of human iPSC-derived RBCs can be an alternative treatment option for patients with blood disorders. In this review, we focused on the generation of human iPSC-derived erythrocytes to present an overview of the current status and applications of this field.

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“…Since hematopoietic cells of the primitive wave are transient and remain of unclear relevance to adult hematopoiesis, cells differentiated from DBA and isogenic iPSCs were harvested between day 19 and 21 (wave 2) of culture for further characterization. differentiation using this approach [31]. The first population peaks early (day 7) and rap-idly disappears.…”

Section: Hematopoietic Differentiation Of Dba Ipscs Phenocopies In VImentioning

confidence: 94%

“…2.11. Hematopoietic Differentiation of DBA and Isogenic iPSCs iPSCs were differentiated for 21 days using the STEMdiff™ Hematopoietic Kit (05310, STEMCELL Technologies, Inc.), as previously described [31]. Briefly, one day be-fore differentiation (Day −1), iPSCs were split as described above, and cluster concentrations were calculated.…”

Section: Identification Of Corrected Ipsc Clonesmentioning

confidence: 99%

“…Protocols primarily based on embryoid body (EB) formation have been utilized in previous studies to facilitate the emergence of human hematopoietic cells from DBA iPSCs but the scale-up of these approaches relies on complex transcription factor-based reprogramming systems to overcome the intrinsic inefficiencies of EB-based iPSC differentiation [ 29 , 30 ]. Here, we applied an alternative differentiation approach to generate erythroid cells from human DBA iPSCs based on a simple, scalable, monolayer platform we previously developed for hematopoietic differentiation of human iPSCs derived from healthy individuals [ 31 ]. Importantly, we find that addition of EPAG during DBA iPSC differentiation significantly improves the erythroid maturation defect recapitulated in this model, suggesting that EPAG may show clinical efficacy in patients with DBA.…”

Section: Introductionmentioning

confidence: 99%

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Eltrombopag Improves Erythroid Differentiation in a Human Induced Pluripotent Stem Cell Model of Diamond Blackfan Anemia

Qanash

Smith

et al. 2021

Cells

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Diamond Blackfan Anemia (DBA) is a congenital macrocytic anemia associated with ribosomal protein haploinsufficiency. Ribosomal dysfunction delays globin synthesis, resulting in excess toxic free heme in erythroid progenitors, early differentiation arrest, and pure red cell aplasia. In this study, DBA induced pluripotent stem cell (iPSC) lines were generated from blood mononuclear cells of DBA patients with inactivating mutations in RPS19 and subjected to hematopoietic differentiation to model disease phenotypes. In vitro differentiated hematopoietic cells were used to investigate whether eltrombopag, an FDA-approved mimetic of thrombopoietin with robust intracellular iron chelating properties, could rescue erythropoiesis in DBA by restricting the labile iron pool (LIP) derived from excessive free heme. DBA iPSCs exhibited RPS19 haploinsufficiency, reduction in the 40S/60S ribosomal subunit ratio and early erythroid differentiation arrest in the absence of eltrombopag, compared to control isogenic iPSCs established by CRISPR/Cas9-mediated correction of the RPS19 point mutation. Notably, differentiation of DBA iPSCs in the presence of eltrombopag markedly improved erythroid maturation. Consistent with a molecular mechanism based on intracellular iron chelation, we observed that deferasirox, a clinically licensed iron chelator able to permeate into cells, also enhanced erythropoiesis in our DBA iPSC model. In contrast, erythroid maturation did not improve substantially in DBA iPSC differentiation cultures supplemented with deferoxamine, a clinically available iron chelator that poorly accesses LIP within cellular compartments. These findings identify eltrombopag as a promising new therapeutic to improve anemia in DBA.

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Robust generation of erythroid and multilineage hematopoietic progenitors from human iPSCs using a scalable monolayer culture system

Cited by 25 publications

References 61 publications

Multifaceted Actions of GFI1 and GFI1B in Hematopoietic Stem Cell Self-Renewal and Lineage Commitment

Multifaceted Actions of GFI1 and GFI1B in Hematopoietic Stem Cell Self-Renewal and Lineage Commitment

Differentiation of human induced pluripotent stem cells into erythroid cells

Eltrombopag Improves Erythroid Differentiation in a Human Induced Pluripotent Stem Cell Model of Diamond Blackfan Anemia

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