Natural Killer and B-Lymphoid Potential in CD34+ Cells Derived From Embryonic Stem Cells Differentiated in the Presence of Vascular Endothelial Growth Factor

Abstract

Differentiation of totipotent mouse embryonic stem (ES) cells to various lymphohematopoietic cells is an in vitro model of the hematopoietic cell development during embryogenesis. To understand this process at cellular levels, differentiation intermediates were investigated. ES cells generated progeny expressing CD34, which was significantly enhanced by vascular endothelial growth factor (VEGF). The isolated CD34+ cells were enriched for myeloid colony-forming cells but not significantly for erythroid colony-forming cells. When cultured on OP9 stroma cells in the presence of interleukin-2 and interleukin-7, the CD34+ cells developed two types of B220+ CD34−lymphocytes: CD3− cytotoxic lymphocytes and CD19+ pre-B cells, and such lymphoid potential was highly enriched in the CD34+ population. Interestingly, the cytotoxic cells expressed the natural killer (NK) cell markers, such as NKR-P1, perforin, and granzymes, classified into two types, one of which showed target specificity of NK cells. Thus, ES cells have potential to generate NK-type cytotoxic lymphocytes in vitro in addition to erythro-myeloid cells and pre-B cells, and both myeloid and lymphoid cells seem to be derived from the CD34+intermediate, on which VEGF may play an important role.

Proteome array identification of bioactive soluble proteins/peptides in Matrigel: relevance to stem cell responses

“…Conversely, PlGF-2 is a positive factor for angiogenesis and endothelial cell proliferation via its binding to the VEGF receptor, and its presence in Matrigel would also be expected to influence Matrigelbased stem cell assays involving blood cell formation and vasculogenesis (Zhou et al 2013). Finally, VEGF itself was detected as a high responder, and again, would mean that Matrigel could, in and of itself, affect ESC hematopoiesis and vasculogenesis, and hematopoietic stem cell differentiation, growth or survival (Nakayama et al 1998;Gerber et al 2002;Gerecht-Nir et al 2003). Chemokines that were indicated to be at high levels in Matrigel by the proteome array results were MIG (145 units) and serpin E1 (160 and 92 units in separate arrays), and some others, 44,and 30 units in separate arrays) and , were detected at lower levels.…”

Evaluation of angiogenesis with vascular endothelial growth factor in patients with thalassemia major

“…In this study, angiogenesis was investigated by measuring serum VEGF in thalassemic patients because VEGF has a prominent role in hematopoietic cell differentiation besides that in normal vascular biology 14–17 . It is suspected that increased bone marrow angiogenesis and VEGF levels are adverse prognostic features in patients with hematological malignancy, such as acute myeloid leukemia or myelodysplastic syndromes 18 .…”

A Novel Chordin-like Protein Inhibitor for Bone Morphogenetic Proteins Expressed Preferentially in Mesenchymal Cell Lineages

“…The E14 ES cells and the OP9 stromal cell line were obtained and cultured as described (Nakayama et al, 1998(Nakayama et al, , 2000. The human kidney epithelial cell lines, 293 and 293T, were maintained in Dulbecco's modified essential medium (DMEM, Gibco/BRL) supplemented with 10% fetal calf serum (FCS, Hyclone, Logan, UT) at 37°C under 5% CO 2 .…”