Figure 1

In vitro screening approach of hematopoiesis-supporting and matrix remodelling capacities of MPN-MSC. (a) Clinical and laboratory characteristics of MPN-MSC donors and control cases. Reticulin fibrosis was evaluated according to the European consensus. y, years; Hb, haemoglobin levels; WBC, white blood cell count; PLT, platelet count; n.d. not determined. (b) BMSCs were isolated from BM of patients with MPN or control donors by plastic adherence. RT-PCR analysis detecting the BCR-ABL gene (left) or JAK2-V617F mutation (right) in cultured BMSC (passage 2). (c) Protein levels of G-CSF and IL-7 in BMSC supernatants (passage 2), (mean ± SD, n = 5 each, *p < 0.05). (d) CFU activity of 500 sorted human CB CD34⁺ cells in the presence of supernatants from BMSC (mean ± SD, n = 5 for control, PCV and CML; n = 4 for ET, *p < 0.05). (e) Contraction of the original collagen gel area (upper panel) in collagen gels after 28 days (mean ± SD, n = 5 each, **p < 0.05). (f) Messenger RNA levels of fibronectin in BMSC in 3D collagen after 28 days of culture. Expression levels are normalized against GAPDH. (mean ± SD, n = 5 each, non-signficant). Quantification of fibronectin expression in collagen gels represented in panel 2–6 by 2 independent researchers (according to grading scale 0–6 below panels). (mean ± SD, n = 5 each, *p < 0.05).