Abstract
Background: The present study investigated the effects of microvascular endothelial cells (MECs) on the chemotaxis, adhesion and proliferation of bone marrow hematopoietic stem cells (HSCs) ex vivo. Methods and Results: MECs were collected from the lung tissue of C57BL/6 mice, and HSCs were isolated with immunomagnetic beads from bone marrow of GFP mice. MECs and HSCs were co-cultured with or without having direct cell–cell contact in Transwell device for the measurement of chemotaxis and adhesion of MECs to HSCs. Experimental results indicate that the penetration rate of HSCs from the Transwell upper chamber to lower chamber in ‘co-culture’ group was significantly higher than that of ‘HSC single culture’ group. Also, the HSCs in co-culture group were all adherent at 24 h, and the co-culture group with direct cell–cell contact had highest proliferation rate. The HSC number was positively correlated with vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1 (SDF-1) levels in supernatants of the culture. Conclusions: Our study reports that MECs enhance the chemotaxis, adhesion and proliferation of HSCs, which might be related to cytokines SDF-1 and VEGF secreted by MECs, and thus MECs enhance the HSC proliferation through cell–cell contact. The present study revealed the effect of MECs on HSCs, and provided a basis and direction for effective expansion of HSCs ex vivo.
Highlights
Hematopoietic stem cell (HSC) is a type of adult stem cells, whose self-renewal, maintenance, apoptosis and motional migration are tightly regulated
They co-cultured Porcine microvascular endothelial cells (PMVEC) and combined cytokines (GM-CSF, IL-3, SCF, IL-6) with purified human CD34 + bone marrow cells in vitro, and observed that in 7 days of culture, the direct contact culture of HSCs with PMVEC monolayers achieved maximum expansion compared to non-contact culture and liquid suspension culture HSCs, and CD34+ hematopoietic stem/progenitor cells were expanded 12.6 times in culture
On the seventh day of cell culture in this experiment, the number of HSCs in the co-culture group (HSCs and microvascular endothelial cells in 2-D) was 41.6 times higher than that in the first day, which was significantly increased compared with Transwell non-contact co-culture group and HSCs culture group
Summary
Hematopoietic stem cell (HSC) is a type of adult stem cells, whose self-renewal, maintenance, apoptosis and motional migration are tightly regulated. It has been reported that ECs provide a nutrient-rich hematopoietic niche for HSC, which can effectively promote HSC proliferation and differentiation [4,5,6,7,8,9,10]. The present study investigated the effects of microvascular endothelial cells (MECs) on the chemotaxis, adhesion and proliferation of bone marrow hematopoietic stem cells (HSCs) ex vivo. MECs and HSCs were co-cultured with or without having direct cell–cell contact in Transwell device for the measurement of chemotaxis and adhesion of MECs to HSCs. Experimental results indicate that the penetration rate of HSCs from the Transwell upper chamber to lower chamber in ‘co-culture’ group was significantly higher than that of ‘HSC single culture’ group. Conclusions: Our study reports that MECs enhance the chemotaxis, adhesion and proliferation of HSCs, which might be related to cytokines SDF-1 and VEGF secreted by MECs, and MECs enhance the HSC proliferation through cell–cell contact. The present study revealed the effect of MECs on HSCs, and provided a basis and direction for effective expansion of HSCs ex vivo
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