Hematopoietic and Endothelial Differentiation of Human Induced Pluripotent Stem Cells

Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, 53715, USA.
Stem Cells (Impact Factor: 6.52). 04/2009; 27(3):559-67. DOI: 10.1634/stemcells.2008-0922
Source: PubMed


Induced pluripotent stem cells (iPSCs) provide an unprecedented opportunity for modeling of human diseases in vitro, as well as for developing novel approaches for regenerative therapy based on immunologically compatible cells. In this study, we employed an OP9 differentiation system to characterize the hematopoietic and endothelial differentiation potential of seven human iPSC lines obtained from human fetal, neonatal, and adult fibroblasts through reprogramming with POU5F1, SOX2, NANOG, and LIN28 and compared it with the differentiation potential of five human embryonic stem cell lines (hESC, H1, H7, H9, H13, and H14). Similar to hESCs, all iPSCs generated CD34(+)CD43(+) hematopoietic progenitors and CD31(+)CD43(-) endothelial cells in coculture with OP9. When cultured in semisolid media in the presence of hematopoietic growth factors, iPSC-derived primitive blood cells formed all types of hematopoietic colonies, including GEMM colony-forming cells. Human induced pluripotent cells (hiPSCs)-derived CD43(+) cells could be separated into the following phenotypically defined subsets of primitive hematopoietic cells: CD43(+)CD235a(+)CD41a(+/-) (erythro-megakaryopoietic), lin(-)CD34(+)CD43(+)CD45(-) (multipotent), and lin(-)CD34(+)CD43(+)CD45(+) (myeloid-skewed) cells. Although we observed some variations in the efficiency of hematopoietic differentiation between different hiPSCs, the pattern of differentiation was very similar in all seven tested lines obtained through reprogramming of human fetal, neonatal, or adult fibroblasts with three or four genes. Although several issues remain to be resolved before iPSC-derived blood cells can be administered to humans for therapeutic purposes, patient-specific iPSCs can already be used for characterization of mechanisms of blood diseases and for identification of molecules that can correct affected genetic networks.

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    • "When total cells were collected from differentiation cultures, the percentage of CD34 + CD45 + cells from different primate species was approximately 20%–30% (Figure 2D). As in humans (Choi et al., 2009a;Vodyanik et al., 2006), the majority of hematopoietic progenitors induced from NHP-iPSCs co-expressed CD43 and CD31 (Figure 2D). Kinetic analysis of differentiation in OP9 cocultures with CHIR99021 reveals striking similarities in hematopoietic differentiation between human iPSCs and NHP-iPSCs. "
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    ABSTRACT: Advances in the scalable production of blood cells from induced pluripotent stem cells (iPSCs) open prospects for the clinical translation of de novo generated blood products, and evoke the need for preclinical evaluation of their efficacy, safety, and immunogenicity in large animal models. Due to substantial similarities with humans, the outcomes of cellular therapies in non-human primate (NHP) models can be readily extrapolated to a clinical setting. However, the use of this model is hampered by relatively low efficiency of blood generation and lack of lymphoid potential in NHP-iPSC differentiation cultures. Here, we generated transgene-free iPSCs from different NHP species and showed the efficient induction of mesoderm, myeloid, and lymphoid cells from these iPSCs using a GSK3β inhibitor. Overall, our studies enable scalable production of hematopoietic progenitors from NHP-iPSCs, and lay the foundation for preclinical testing of iPSC-based therapies for blood and immune system diseases in an NHP model.
    Full-text · Article · Feb 2016 · Stem Cell Reports
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    • "Induced pluripotent stem cells (iPSCs) are a novel cell type, derived from somatic cell reprogramming via overexpression of exogenous transcription factors [3]. These reprogrammed pluripotent cells are then capable of being differentiated into many different mature cell types, including vascular endothelial cells (iPSC-ECs) [4] [5] [6]. Similarly, induced endothelial cells (iECs) can be produced by transdifferentiating adult fibroblasts directly to endothelial cells, bypassing the pluripotent stem cell intermediate [7] [8]. "

    Full-text · Article · Jan 2015
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    • "Once the cells have returned to a pluripotent state the cells can then be re-differentiated into endothelial and associated mural cells. Choi et al. were able to differentiate seven human iPS cell lines into endothelial (CD31+, CD34-) cells [61]. The iPS-derived endothelial cells were shown to successfully form capillary-like structures on growth factor reduced matrigel in 2D. "
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    ABSTRACT: In vitro pre-vascularization is one of the main vascularization strategies in the tissue engineering field. Culturing cells within a tissue-engineered construct (TEC) prior to implantation provides researchers with a greater degree of control over the fate of the cells. However, balancing the diverse range of different cell culture parameters in vitro is seldom easy and in most cases, especially in highly vascularized tissues, more than one cell type will reside within the cell culture system. Culturing multiple cell types in the same construct presents its own unique challenges and pitfalls. The following review examines endothelial-driven vascularization and evaluates the direct and indirect role other cell types have in vessel and capillary formation. The article then analyses the different parameters researchers can modulate in a co-culture system in order to design optimal tissue-engineered constructs to match desired clinical applications.
    Full-text · Article · Jun 2014 · Vascular Cell
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