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In vitro Cultivation of MesenchymalStem Cells in Patients with Lymphoid Malignancies


Authors: E. Novotová 1;  H. Strnadová 1;  B. Procházka 2;  R. Pytlík 1
Authors‘ workplace: 1. interní klinika 1. LF UK a VFN, Praha 2Oddělení biomedicíny a statistiky, Státní zdravotní ústav, Praha 1
Published in: Transfuze Hematol. dnes,, 2003, No. 1, p. 30-36.
Category:

Overview

Background.
Mesenchymal stem cells (MSC) are currently extensively studied because of their abilitiesto support haemopoiesis, to modulate immune reactions (including GVHD) and to differentiate intospecialized tissues (cartilage, bone, smooth muscle). In our laboratory we studied growth properties ofmesenchymal stem cells in patients with lymphoid malignancies. Materials and methods. 10 ml of bonemarrow aspirate was taken from patients undergoing routine diagnostic or control trephine biopsy.After separation of mononuclear cells (MNC), adhesive MNCs and fibroblasts were counted and theirnumber expressed per 105 total MNCs. Mesenchymal cells were grown in complete medium (α-MEM +20% fetal bovine serum + glutamine + antibiotics). We evaluated the number of colonies from the 1stpassage (per 105 total MNCs = CFU-F1), number of MSCs from the 1st passage and number of MSCcolonies grown from the 2nd passage (CFU-F2) after seeding in concentrations of 1.5, 3, 5 a 10 cells/cm2.Results. Samples were taken from 44 patients; from 15 of them bilateral samples were obtained. Thenumber of CFU-F1 correlated significantly only with the number of adhesive fibroblasts. The numberof CFU-F2 colonies showed an approximately linear correlation with the concentrations of cells seeded.Patients with chronic lymphocytic leukaemia and multiple myeloma had higher CFU-F2 counts thanpatientswith lymphoma.When bilateral samples were taken, very good correlations between the countsof adherent MNCs, fibroblasts, CFU-F1 and CFU-F2 were observed in both samples from the samepatient. Conclusion. Our method of evaluation of MSC numbers in bone marrow seems reproducibleand reliable. Different growth characteristics of MSCs in patients with different lymphoidmalignanciescan be caused by different profiles of cytokine secretion, which can in turn influence the pathogenesisof these diseases.

Key words:
mesenchymal stem cells, lymphoma, chronic lymphocytic leukaemia, myeloma

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Haematology Internal medicine Clinical oncology
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