How the niche regulates hematopoietic stem cells.

The hematopoietic stem cell (HSC) forms all types of blood cells of the hematopoietic system. In the adult, HSC are mainly quiescent, being mostly in G0/G1 phase of cell cycle during steady-state conditions. However, during hematopoietic stress, the stem cells respond quickly to regenerate the damaged hematopoietic system. To understand how environmental signals affect HSC and its progeny, it is essential to know the lineage relationships and transcriptional mechanisms controlling self-renewal, proliferation and differentiation. Because of the high possible output of blood cells from a single HSC, a tight regulation of these processes is extremely important. An essential component for this control is the marrow microenvironment, in this context also referred to as the HSC niche. The niche is heterogeneous and regulates stem cell metabolism through both surface-bound and soluble factors. Several signaling pathways have been shown to take part in these regulation processes, with Notch and especially Wnt signaling being the best studied ones. Dysregulation of the niche, for instance by environmental exposure, has recently been shown to lead to hematopoietic abnormalities. Thus, to understand the effect of the environment on hematopoiesis, it is of importance to study both HSC, its direct progeny and the cellular components of the niche. Detailed knowledge of the regulatory mechanisms operating between hematopoietic cells and their direct surroundings facilitates the study of how such signaling may be disrupted by environmental exposure.

Secreted frizzled-related protein 1 extrinsically regulates cycling activity and maintenance of hematopoietic stem cells.

Secreted frizzled-related protein 1 (Sfrp1) is highly expressed by stromal cells maintaining hematopoietic stem cells (HSCs). Sfrp1 loss in stromal cells increases production of hematopoietic progenitors, and in knockout mice, dysregulates hemostasis and increases Flk2- Cd34- Lin- Sca1+ Kit+ (LSK) cell numbers in bone marrow. Also, LSK and multipotent progenitors (MPPs) resided mainly in the G0/G1 phase of cell cycle, with an accompanying decrease in intracellular beta-catenin levels. Gene-expression studies showed a concomitant decrease Ccnd1 and Dkk1 in Cd34- LSK cells and increased expression of Pparg, Hes1, and Runx1 in MPP. Transplantation experiments showed no intrinsic effect of Sfrp1 loss on the number of HSCs or their ability to engraft irradiated recipients. In contrast, serial transplantations of wild-type HSCs into Sfrp1(-/-) mice show a progressive decrease of wild-type LSK and MPP numbers. Our results demonstrate that Sfrp1 is required to maintain HSC homeostasis through extrinsic regulation of beta-catenin.

Efficient hematopoietic differentiation of human embryonic stem cells on stromal cells derived from hematopoietic niches.

Hematopoietic stem cells derived from human embryonic stem cells (hESCs) could provide a therapeutic alternative to bone marrow transplants, but the efficiency of currently available derivation protocols is low. In this study, we investigated whether coculture with monolayers of cells derived from mouse AGM and fetal liver, or with stromal cell lines derived from these tissues, can enhance hESC hematopoietic differentiation. We found that under such conditions hESC-derived differentiating cells formed early hematopoietic progenitors, with a peak at day 18-21 of differentiation that corresponded to the highest CD34 expression. These hESC-derived hematopoietic cells were capable of primary and secondary hematopoietic engraftment into immunocompromised mice at substantially higher levels than described previously. Transcriptional and functional analysis identified TGF-beta1 and TGF-beta3 as positive enhancers of hESC hematopoietic differentiation that can further stimulate this process when added to the culture. Overall, our findings represent significant progress toward the goal of deriving functional hematopoietic stem cells from hESCs.