In vitro model of myelofibrosis using human platelet lysate

EXPERIMENTAL STUDIES , , ,

Ye.N. Bulycheva, N.T. Siordiya, E.G. Lomaia, A.Yu. Zaritskiy, and P.A. Butylin

V.A. Almazov Federal Heart, Blood and Endocrinology Center, Saint Petersburg, Russian Federation

ABSTRACT

The development and studies of the myelofibrosis (MF) in vitro model is an important issue, since such model can lead to understanding of pathogenesis and identifying the new targets for therapy.

Objectives. Here, we studied the properties of mesenchymal stromal cells (MSCs) cultured in the medium containing the human platelet lysate (HPL).

Design and methods. Bone marrow MSCs from healthy donors and a patient with primary myelofibrosis (PMF) were cultured in the media containing various HPL concentrations. We measured the proliferative activity, the collagen type I and III expression, and capability to differentiate into the osteogenic or adipogenic lineages. The concentrations of the vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), transforming growth factor beta (TGF-b), and hepatocyte growth factor (HGF) were measured in HPL from 17 patients with primary myelofibrosis or post-polycythemia vera myelofibrosis (post-PV MF) using the specific ELISA kits.

Results. The highest MSC proliferative rate was found in the cultures with high HPL concentrations (10–20 %). The ratio of collagen type III/collagen type I expression was the highest in the cultures containing 10 % HPL. The use of HPL for MSCs culturing caused no changes in their osteogenic differentiation capability, but the increase in the HPL concentration resulted in the decreased capability to differentiate into the adipogenic lineage. Further, we observed the significantly increased VEGF and bFGF concentrations in HPL from MF patients, compared to the age-matched healthy controls (2.5- and 2.4-fold, respectively, p < 0.01), while the TGF-b and HGF concentrations showed the trend towards an increase, but with no significant difference from the controls. MSCs cultured with HPL from MF patients showed a higher proliferative rate compared to HPL from healthy donors. MSCs from the PMF patient tended to proliferate more actively compared to the cells from healthy donors.

Conclusion. MSCs culturing using varying HPL concentrations can be used as an adequate MF in vitro model, since it leads to pro-fibrotic changes in the bone marrow stromal cells.

Keywords: primary myelofibrosis, platelet lysate, mesenchymal stromal cells, myelofibrosis in vitro model.

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