Megakaryocyte?stromal cell interactions: Effect on megakaryocyte proliferation, proplatelet production, and survival

NORA PAULA GOETTE; FRANCISCO R. BORZONE; AILEN D. DISCIANNI LUPI; NORMA A. CHASSEING; MARIA F. RUBIO; MONICA A. COSTAS; PAULA G. HELLER; ROSANA F. MARTA; PAOLA R. LEV

EXPERIMENTAL HEMATOLOGY

ELSEVIER SCIENCE INC

Lugar: Amsterdam; Año: 2022 vol. 107 p. 24 - 37

0301-472X

Bone marrow stromal cells provide a proper environment for thedevelopment of hematologic lineages. The incorporation of differentstromal cells into in vitro culture systems would be an attractivemodel to study megakaryopoiesis and thrombopoiesis. Our objectivewas to evaluate the participation of different types of stromal cells inin vitro megakaryopoiesis, thrombopoiesis, and megakaryocyte (MK)survival. CD34-positive progenitors from umbilical cord blood weredifferentiated into MK precursors and then cocultured with umbilicalvein endothelial cells (HUVECs), bone marrow mesenchymal stemcells (MSCs), skin fibroblasts (SFs) (all human), or the mouse fibroblastcell line L929. The number of MKs (CD61-positive cells) wasincreased in the presence of HUVECs and SFs, whereas L929 cellsdecreased total and mature MK counts. With respect to thrombopoiesis,HUVECs increased proplatelet (PP)-producing MKs, whileMSCs, L929 cells, and SFs had the opposite effect (immunofluorescencestaining and microscopic analysis). MK survival was enhancedin MSC and SF co-cultures, as assessed by evaluation of pyknoticnuclei. However, HUVECs and L929 did not prevent apoptosis ofMKs. Reciprocally, the cloning efficiency of MSCs was decreased inthe presence of MKs, while the ability of stromal cells (either MSCsor SFs) to produce the extracellular matrix proteins type III collagen,fibronectin, dermatan sulfate, heparan sulfate, and prolyl 4-hydroxylasesubunit b was preserved. These data indicate that each stromalcell type performs distinctive functions that differentially modulateMK growth and platelet production and, at the same time, that MKsalso modify stromal cell behavior. Overall, our results highlight the relevanceof considering the influence of stromal cells in MK research aswell as the close interplay of different cell types within the bone marrowmilieu.