Article

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: 7.7). 04/2009; 27(3):559-67. DOI: 10.1634/stemcells.2008-0922
Source: PubMed

ABSTRACT 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.

0 Bookmarks
 · 
138 Views
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Chronic granulomatous disease (CGD) is an inherited orphan disorder caused by mutations in one of the five genes encoding reduced nicotinamide-adenine-dinucleotide-phosphate oxidase subunits, which subsequently lead to impairment in the production of microbicidal reactive oxygen species (ROS). In order to offer several cell line models of CGD and therefore support research on pathophysiology and new therapeutic approaches, we optimized protocols to differentiate induced pluripotent stem cells (iPSCs) from wild-type, X 0 -, AR22 0 -and AR47 0 -CGD patient's fibroblasts into neutrophils and into macrophages. Aberrant genetic clones were dis-carded after chromosome karyotyping and array-comparative genomic hybridization analysis. All remaining iPSC lines showed human embryonic stem cell–like morphology, expressed all tested pluripotency markers and formed embryoid bodies that contained cells originating from all three primary germ layers. Furthermore, each CGD patient-specific iPSC line retained the gp91 phox , p47 phox , and p22 phox mutations found in the corre-sponding patient's neutrophils. The average production of CD34 + progenitors was of 1.5 · 10 6 cells after 10 days of differentiation of 10 · 10 6 iPSCs. They were terminally differentiated into about 3 · 10 5 neutrophils or into 3 · 10 7 macrophages. Based on morphological, phenotypical, and functional criteria both phagocyte types were mature and indistinguishable from the native human neutrophils and macrophages. However, neutrophils and macrophages derived from X 0 -, AR22 0 -, and AR47 0 -CGD patient-specific iPSC lines lacked ROS produc-tion and the corresponding mutated proteins. To simplify the phagocytes' production upon request, progenitors can be cryopreserved. In conclusion, we describe a reproducible, simple, and efficient way to generate neutro-phils and macrophages from iPSCs and provide a new cellular model for the AR22 0 -CGD genetic form that has not been described before.
    BioResearch Open Access. 12/2014;
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: A major challenge modern society has to face is the increasing need for tissue regeneration due to degenerative diseases or tumors, but also accidents or warlike conflicts. There is great hope that stem cell-based therapies might improve current treatments of cardiovascular diseases, osteochondral defects or nerve injury due to the unique properties of stem cells such as their self-renewal and differentiation potential. Since embryonic stem cells raise severe ethical concerns and are prone to teratoma formation, adult stem cells are still in the focus of research. Emphasis is placed on cellular signaling within these cells and in between them for a better understanding of the complex processes regulating stem cell fate. One of the oldest signaling systems is based on nucleotides as ligands for purinergic receptors playing an important role in a huge variety of cellular processes such as proliferation, migration and differentiation. Besides their natural ligands, several artificial agonists and antagonists have been identified for P1 and P2 receptors and are already used as drugs. This review outlines purinergic receptor expression and signaling in stem cells metabolism. We will briefly describe current findings in embryonic and induced pluripotent stem cells as well as in cancer-, hematopoietic-, and neural crest-derived stem cells. The major focus will be placed on recent findings of purinergic signaling in mesenchymal stem cells addressed in in vitro and in vivo studies, since stem cell fate might be manipulated by this system guiding differentiation towards the desired lineage in the future.
    Computational and Structural Biotechnology Journal. 11/2014;
  • [Show abstract] [Hide abstract]
    ABSTRACT: The integral moment method based on the curved ray-tracing method is proposed to solve radiative transfer in a one-dimensional nonlinear anisotropic scattering medium with graded index. This method does not suffer from ray effects since there is no angular discretization. A steady-state combined conduction and radiation heat transfer problem is considered. Temperature and radiative heat flux distributions are determined through the spatial discretization. The results show that the integral moment method is good in accuracy and convergence characteristic. Various effects of nonlinear anisotropic scattering and graded index on radiative transfer are investigated.
    International Journal of Thermal Sciences 09/2012; 59:58-65. · 2.56 Impact Factor

Preview

Download
2 Downloads
Available from