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

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, find- IntroductionHuman embryonic stem cells (hESCs) provide an important model system to define the mechanisms that mediate cellular development. hESC-derived hematopoietic progenitor cells efficiently produce erythroid, myeloid, and lymphoid lineage cells in vitro. [1][2][3][4] We previously defined an in vitro culture system to generate natural killer (NK) cells from hESCs. 5 hESC-derived NK cells express surface receptors characteristic of primary NK cells, kill tumor target cells, and produce interferon-␥ when stimulated with cytokines. These results suggest that hESC-derived progenitors may also readily commit to the T-cell lineage in vitro, since T and NK lymphocytes are developmentally closely related. 6,7 One study has used an in vivo model to examine the T-cell potential of hESCs. 8 Galic et al injected hESC-derived hematopoietic progenitor cells into human thymus/fetal liver (Thy/Liv) grafts in severe combined immunodeficient-human (SCID-hu) mice. This study demonstrated T-cell development after 3 to 5 weeks in vivo, although in a less efficient manner than what has been observed with hematopoietic progenitor cells from human fetal liver (FL), bone marrow (BM), or umbilical cord blood (UCB) 9-11 evaluated in SCID-hu mice. Although useful, SCID-hu mice are not optimal to evaluate development of specific phenotypic cell populations over time, and the effects of specific molecular signaling pathways are difficult to quantify via this SCID-hu system. Therefore, in vitro models of lymphocyte development are needed, although despite the considerable interest in hematopoietic development of hESCs, in vitro studies have not provided significant evidence of functional T and B lymphocyte maturation of hESC-derived hematopoietic progenitors. Al...

Mechanisms of thymus organogenesis and morphogenesis

“…A report of anterior foregut endoderm derived from human ES and IPS cells holds more promise for robust and controlled generation of TECs, although that goal has not yet been reached . By contrast, substantial progress has been reported in generating T cells from both human and mouse hematopoietic stem cells or from ES cells in vitro (Schmitt et al, 2004;de Pooter and Zuniga-Pflucker, 2007;Awong et al, 2008;Martin et al, 2008) using an OP9 cell line expressing delta-like 1 (OP9-DL1) that can sustain most aspects of T cell differentiation in vitro (Schmitt and Zuniga-Pflucker, 2002;Holmes and ZunigaPflucker, 2009). So, why make TECs at all?…”

Generation of T Cells from Human Embryonic Stem Cell-Derived Hematopoietic Zones

“…Although T precursor activity was not addressed, these data suggest that the CD34 ϩ CD43 ϩ population may contain multipotent HPC. However, a recent report from Martin et al (7) indicated that such hESC-derived HPC lack T lineage potential. hESC-derived CD34 ϩ cells, introduced in fetal thymic organ cultures or in cocultures with Delta-like 1 expressing OP9 stromal cells (OP9-DL1), did not generate T cells, in contrast to cord blood-derived CD34 ϩ cells.…”

Generation of Blood Cells from Human Embryonic Stem Cells and Their Possible Clinical Utilization

“…To allow more access to developing cells and improving conditions that support or inhibit development of T cells, a Notch ligand-expressing OP9-DL1 stromal cells have been used to derive T cells from multiple progenitor cell populations expressing CD34 and CD45 such as human BM, umbilical cord blood, and mouse ESCs (RF. de Pooter , et al 2003;CH. Martin, et al, 2008;TM.…”