Expansion of human hematopoietic stem/progenitor cells on decellularized matrix scaffolds

Umbilical cord blood (UCB) is one of the richest sources for hematopoietic stem/progenitor cells (HSPCs), with more than 3000 transplantations performed each year for the treatment of leukemia and other bone marrow, immunological, and hereditary diseases. However, transplantation of single cord blood units is mostly restricted to children, due to the limited number of HSPC per unit. This unit develops a method to increase the number of HSPCs in laboratory conditions by using cell-free matrices from bone marrow cells that mimic 'human-body niche-like' conditions as biological scaffolds to support the ex vivo expansion of HSPCs. In this unit, we describe protocols for the isolation and characterization of HSPCs from UCB and their serum-free expansion on decellularized matrices. This method may also help to provide understanding of the biochemical organization of hematopoietic niches and lead to suggestions regarding the design of tissue engineering-based biomimetic scaffolds for HSPC expansion for clinical applications.

Heterogeneous copper concentrations in cancerous human prostate tissues

© 2015 Wiley Periodicals, Inc. © 2015 Wiley Periodicals, Inc. Background Therapeutics that target copper for the treatment of prostate cancer are being evaluated in human clinical trials. Elevated intracellular copper is considered to sensitize prostate cancer cells to certain copper-coordination compounds, especially those with ionophoric properties. While there is compelling in vitro evidence that prostate cancer cells accumulate intracellular copper, a corresponding status for copper in patient tissues has not been corroborated. We therefore established whether copper concentrations increase in cancerous prostate tissues, and in sera, in patients throughout disease progression. Methods Human prostate tissue samples were obtained from patient prostatectomies (n=28), and together with patient-matched sera, were analyzed for copper content by inductively coupled plasma mass spectrometry. Results When grouped together, cancerous prostate tissues exhibiting moderate disease severity (Gleason Score 7) (n=10) had 1.6-fold more copper than age-matched normal tissues (n=10) (P<0.05). Those with more aggressive disease (Gleason Score 9) (n=8) had 1.8-fold more copper (P<0.05). In both disease stages however, the copper concentrations between individual samples were rather variable (0.55-3.02μg/g), with many clearly within the normal range (0.52-1.28μg/g). Additionally, we found that there was no change in serum copper concentrations in patients with either moderate or aggressive prostate cancer (Gleason Score 7 or 9), compared with reference intervals and to age-matched controls. Conclusions The heterogeneous nature of copper concentrations in cancerous prostate tissues, suggest that a small subset of patients may respond to treatments that target elevated intratumoral copper. Therefore, such approaches would likely require personalized treatment strategies. Prostate 75:1510-1517, 2015.