Erythroid differentiation of human induced pluripotent stem cells is independent of donor cell type of origin

Dorn, Isabel; Klich, Katharina; Araúzo‐Bravo, Marcos J.; Radstaak, Martina; Santourlidis, Simeon; Ghanjati, Foued; Radke, Teja Falk; Psathaki, Olympia E.; Hargus, Gunnar; Kramer, Jan; Einhaus, M; Kim, Jeong Beom; Kögler, Gesine; Wernet, Peter; Schöler, Hans R.; Schlenke, Peter; Zaehres, Holm

doi:10.3324/haematol.2014.108068

Cited by 80 publications

(100 citation statements)

References 56 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…2A) [20]. The generation of a Expression of surface markers specific for hematopoietic specification (KDR (VEGFR2), CD144, CD41a, CD235a (GPA), CD43) was assessed on EB day 10, 15 and 21 by flow cytometry (B-G).…”

Section: Hematopoietic and Erythroid Differentiation Of Hescs And Hipscsmentioning

confidence: 99%

“…CB-derived iPSC line by OCT4, SOX2, KLF4, and c-Myc and its characterization has been described previously [20]. CD43+ cells as well as CD43 low and CD43 hi subpopulations were seeded at a density of 5 × 10 4 cells/ml upon in vitro erythropoiesis initiation (stage II).…”

Section: Hematopoietic and Erythroid Differentiation Of Hescs And Hipscsmentioning

confidence: 99%

“…Here, we generated CD43-expressing cells using a xeno-product-and cell free embryoid body(EB)-based differentiation protocol [20]. We investigated the erythroid differentiation potential of CD43 expressing cells and defined a CD43 hi subset as the source of erythroid differentiation from hiPSCs.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Emergence of CD43-Expressing Hematopoietic Progenitors from Human Induced Pluripotent Stem Cells

Kessel¹,

Bluemke²,

Schöler³

et al. 2017

Transfus Med Hemother

View full text Add to dashboard Cite

Background: The ex vivo generation of human hematopoietic stem cells (HSCs) with long-term repopulating capacity and multi-lineage differentiation potential represents the holy grail of hematopoiesis research. In principle, human induced pluripotent stem cells (hiPSCs) provide the tool for both studying molecular mechanisms of hematopoietic development and the ex vivo production of ‘true' HSCs for transplantation purposes and lineage-specific cells, e.g. red blood cells, for transfusion purposes. CD43-expressing cells have been reported as the first hematopoietic cells during development, but whether or not these possess multilineage differentiation and long-term engraftment potential is incompletely understood. Methods: We performed ex vivo generation of hematopoietic cells from hiPSCs using an embryoid body(EB)-based, xeno-product-free differentiation protocol. We investigated the multilineage differentiation potential of different FACS-sorted CD43-expressing cell subsets by colony-forming assays in semisolid media. Further, erythroid differentiation was investigated in more detail using established protocols. Results: By using CD43, we were able to measure hematopoietic induction efficiency during hiPSC-derived EB differentiation. Further, we determined CD43+ cells as the cell population of origin for in vitro erythropoiesis. Furthermore, colony formation demonstrates that the multipotent hematopoietic stem and progenitor cell fraction is particulary enriched in the CD43^hi CD45+ population.

show abstract

Section: Hematopoietic and Erythroid Differentiation Of Hescs And Hipscsmentioning

confidence: 99%

Section: Hematopoietic and Erythroid Differentiation Of Hescs And Hipscsmentioning

confidence: 99%

See 1 more Smart Citation

Emergence of CD43-Expressing Hematopoietic Progenitors from Human Induced Pluripotent Stem Cells

Kessel¹,

Bluemke²,

Schöler³

et al. 2017

Transfus Med Hemother

View full text Add to dashboard Cite

show abstract

“…Disease modelling of beta haemoglobinopathies remains unattained as erythroid cells obtained via direct differentiation of iPSCs retain an immature phenotype with little expression of the -globin gene [148][149][150][151][152]154]. This study sought to assess the applicability of CRISPRa for stimulating gene expression of the -globin locus inactive in iPSCs, explore the potential of this system for modelling the RNA processing defect in beta thalassaemia and validate the restoration of functionality of the wild type allele in gene-corrected iPSCs.…”

Section: Discussion and Future Directionsmentioning

confidence: 99%

“…Indeed, the -globin gene is only expressed during the late stages of erythropoiesis and is considered an adult attribute (HbA). It has been shown that erythroid cells, derived by the currently available methods for directed in vitro differentiation of iPSCs, mostly adopt an embryonic phenotype with variable expression of foetal globin and failure of efficient globin switching to -globin [154]. To circumvent this problem, in this study an alternative approach was considered, which takes advantage of CRISPR-mediated transcriptional activation of the endogenous gene [156] to assess the restoration of adult gene expression following gene correction of a mutation associated with a loss of function in monogenic disease.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9-mediated traceless gene correction and activation of the HBB locus in iPSCs with the beta thalassaemia mutation

Al-Ateeq¹

View full text Add to dashboard Cite

Beta thalassaemia is caused by mutations in the adult haemoglobin gene (HBB) and is one of the most prevalent monogenic blood disorders worldwide. The transplantation of haematopoietic stem cells derived from gene-corrected patient induced pluripotent stem cells (iPSCs) could provide a promising therapeutic strategy. Here, the generation and characterisation of footprint-free iPSCs from dermal fibroblasts of an individual with a homozygous beta thalassaemia-causing mutation affecting splicing is described (Chapter III). Successful gene correction of the disease-causing mutation was achieved by employing a CRISPR/Cas9 dual nickase approach and two donor design strategies aimed at reducing undesired on-target mutagenesis (Chapter IV), followed by the This study combined cutting-edge cellular reprogramming methods to generate beta thalassaemia iPSCs, a double nickase-mediated gene editing approach and piggyBac transposase-aided excision to achieve seamless gene repair, and a CRISPRa approach to model the impact of the diseasecausing mutation and demonstrate restoration of normal transcription following genetic repair.Having established a platform for precise gene correction and for validation of the restoration of the functional allele in this monogenic disease, this strategy may provide a viable avenue for iPSCiii based cell therapy of beta thalassaemic patients provided mature engraftable blood cells can be generated from hiPSCs in the future.iv

show abstract

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

et al. 2019

View full text Add to dashboard Cite

In vitro cultured blood cells for transfusion purposes provide a safe alternative to donor blood, particularly for patients who require recurrent transfusions, and can be used as carriers of therapeutic molecules. In vitro derivation of hematopoietic cell types from human‐induced pluripotent stem cells (iPSCs) allows for a constant, well‐defined production pipeline for such advanced therapeutic and medicinal products. Application of selected iPSC‐derived hematopoietic stem cells and hematopoietic effector cells in transplantation/transfusions would avoid the risk of alloimmunization and blood‐borne diseases, as well as enable the production of enhanced blood cells expressing molecules that enforce blood cell function or endow novel therapeutic properties. Here, we discuss the state of the art approaches to produce erythroid, megakaryoid and myeloid cells from iPSCs and the biological and technical hurdles that we need to overcome prior to therapeutic application.

show abstract

Erythroid differentiation of human induced pluripotent stem cells is independent of donor cell type of origin

Cited by 80 publications

References 56 publications

Emergence of CD43-Expressing Hematopoietic Progenitors from Human Induced Pluripotent Stem Cells

Emergence of CD43-Expressing Hematopoietic Progenitors from Human Induced Pluripotent Stem Cells

CRISPR/Cas9-mediated traceless gene correction and activation of the HBB locus in iPSCs with the beta thalassaemia mutation

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

Contact Info

Product

Resources

About