Differences in lymphocyte developmental potential between human embryonic stem cell and umbilical cord blood-derived hematopoietic progenitor cells

Stem Cell Institute, Department of Medicine, University of Minnesota, Minneapolis, USA.
Blood (Impact Factor: 10.43). 08/2008; 112(7):2730-7. DOI: 10.1182/blood-2008-01-133801
Source: PubMed

ABSTRACT 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, Aug 12, 2015
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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
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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 embryonic stem cells (hESC) have the potential to revolutionize certain medical treatments, including T-cell-based therapies. However, optimal approaches to develop T cells from hESC are lacking. In this report, we show that T-cell progenitors can be derived from hESC cultured as embryoid bodies (EBs). These EB-derived T-cell progenitors give rise to phenotypically and functionally normal cells of the T lineage when transferred into human thymic tissue implanted in immunocompromised mice, suggesting that introduction of these progenitors into patients may also yield functional T cells. Moreover, hematopoietic progenitors demonstrating T-cell potential appeared to be CD45+/CD34+, resembling those found in normal bone marrow. In contrast to T cells developed from hESC cocultured on murine stromal cells, the EB-derived T cells also expressed normal levels of CD45. Importantly, the EB system eliminates the previous need for murine cocultures, a key impediment to developing a protocol for T-cell progenitor derivation suitable for clinical use. Furthermore, following lentiviral-mediated introduction of a vector expressing enhanced green fluorescent protein into hESC, stable transgene expression was maintained throughout differentiation, suggesting a potential for gene therapy approaches aimed at the augmentation of T-cell function or treatment of T-cell disorders.
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