Martin, CH, Woll, PS, Ni, Z, Zúñiga-Pflücker, JC and Kaufman, DS. Differences in lymphocyte developmental potential between human embryonic stem cell and umbilical cord blood-derived hematopoietic progenitor cells. Blood 112: 2730-2737

Stem Cell Institute, Department of Medicine, University of Minnesota, Minneapolis, USA.
Blood (Impact Factor: 10.45). 08/2008; 112(7):2730-7. DOI: 10.1182/blood-2008-01-133801
Source: PubMed


Hematopoietic progenitor cells derived from human embryonic stem cells (hESCs) develop into diverse mature hematopoietic lineages, including lymphocytes. Whereas functional natural killer (NK) cells can be efficiently generated in vitro from hESC-derived CD34(+) cells, studies of T- and B-cell development from hESCs have been much more limited. Here, we demonstrate that despite expressing functional Notch-1, CD34(+) cells from hESCs did not derive T cells when cocultured with OP9 cells expressing Delta-like 1, or in fetal thymus organ culture. hESC-derived CD34(+) cells also did not produce B cells in vitro. In contrast, CD34(+) cells isolated from UCB routinely generated T and B cells when cultured in the same conditions. Notably, both undifferentiated hESCs, and sorted hESC-derived populations with hematopoietic developmental potential exhibited constitutive expression of ID family genes and of transcriptional targets of stem cell factor-induced signaling. These pathways both inhibit T-cell development and promote NK-cell development. Together, these results demonstrate fundamental differences between hESC-derived hematopoietic progenitors and analogous primary human cells. Therefore, hESCs can be more readily supported to differentiate into certain cell types than others, findings that have important implications for derivation of defined lineage-committed populations from hESCs.

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Available from: Juan Carlos Zúñiga-Pflücker
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    • "EBs obtained from pluripotent stem cells were differentiated on OP9 (Martin et al., 2008) and OP9-Delta1 (OP9D1) (Kennedy et al., 2012) stroma monolayer for NK cell and T cell differentiation, respectively, in Dulbecco's modified Eagle's medium supplemented with 20% fetal bovine serum. To obtain NK cells, EBs were differentiated for 4 weeks in the presence of interleukin-3 (IL-3) (5 ng/ml), IL-7 (5 ng/ml), stem cell factor (SCF) (10 ng/ml), and Flt-3L (10 ng/ml) (all from PeproTech). "
    Dataset: 2015-Ronn

    Full-text · Dataset · Apr 2015
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    • "Furthermore, we show that across iPSC lines, erythroblasts lack expression of Class I HLA, which mirrors normal erythropoiesis, and has important implications both for diagnostics and transfusion medicine. Given that there is still much controversy regarding the derivation of B cells from pluripotent stem cells (Martin et al, 2008), it is of note that B cell lymphopoiesis was demonstrated from multiple hiPSCs sources. Additionally, we have described various somatic and acquired mutations across hiPSCs lines, where some CNVs are common in hiPSC lines, as described previously (Martins-Taylor et al, 2011). "
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    ABSTRACT: Human induced pluripotent stem cells (hiPSCs), like embryonic stem cells, are under intense investigation for novel approaches to model disease and for regenerative therapies. Here, we describe the derivation and characterization of hiPSCs from a variety of sources and show that, irrespective of origin or method of reprogramming, hiPSCs can be differentiated on OP9 stroma towards a multi-lineage haemo-endothelial progenitor that can contribute to CD144+ endothelium, CD235a+ erythrocytes (myeloid lineage) and CD19+ B lymphocytes (lymphoid lineage). Within the erythroblast lineage, we were able to demonstrate by single cell analysis (flow cytometry), that hiPSC-derived erythroblasts express alpha globin as previously described, and that a sub-population of these erythroblasts also express haemoglobin F (HbF), indicative of fetal definitive erythropoiesis. More notably however, we were able to demonstrate that a small sub-fraction of HbF positive erythroblasts co-expressed HbA in a highly heterogeneous manner, but analogous to cord blood-derived erythroblasts when cultured using similar methods. Moreover, the HbA expressing erythroblast population could be greatly enhanced (44·0 ± 6·04%) when a defined serum-free approach was employed to isolate a CD31+ CD45+ erythro-myeloid progenitor. These findings demonstrate that hiPSCs may represent a useful alternative to standard sources of erythrocytes (RBCs) for future applications in transfusion medicine.
    Full-text · Article · May 2014 · British Journal of Haematology
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    • "The comparatively high expression of Id factors in hESCs and hESC-derived hematopoietic cells as compared to cord blood hematopoietic progenitors has been noted. Id genes antagonize T lineage development and may be one of the hurdles to in vitro T cell generation from hESC lines [15]. An additional test of T lineage potential may be passage of hESC-derived hematopoietic progenitors through a humanized mouse model. "
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    ABSTRACT: Directed differentiation of human embryonic stem cells (hESCs) into any desired cell type has been hailed as a therapeutic promise to cure many human diseases. However, substantial roadblocks still exist for in vitro differentiation of hESCs into distinct cell types, including T lymphocytes. Here we examined the hematopoietic differentiation potential of six different hESC lines. We compare their ability to develop into CD34(+) or CD34(+)CD45(+) hematopoietic precursor populations under several differentiation conditions. Comparison of lymphoid potential of hESC derived- and fetal tissue derived-hematopoietic precursors was also made. We found diverse hematopoietic potential between hESC lines depending on the culture or passage conditions. In contrast to fetal-derived hematopoietic precursors, none of the CD34(+) precursors differentiated from hESCs were able to develop further into T cells. These data underscore the difficulties in the current strategy of hESC forward differentiation and highlight distinct differences between CD34(+) hematopoietic precursors generated in vitro versus in vivo.
    Full-text · Article · May 2011 · PLoS ONE
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