title: Growth Factor Content in Human Sera Affects the Isolation of Mesangiogenic Progenitor Cells (MPCs) from Human Bone Marrow.
abstract: Mesangiogenic Progenitor Cells (MPCs) are human bone marrow-derived multipotent cells, isolatedin vitrounder selective culture conditions and shown to retain both mesengenic and angiogenic potential. MPCs also co-isolated with multipotent stromal cells (MSCs) when bone marrow primary cultures were set up for clinical applications, using human serum (HS) in place of fetal bovine serum (FBS). MPC culture purity (over 95%) is strictly dependent on HS supplementation with significant batch-to-batch variability. In the present paper we screened different sources of commercially available pooled human AB type serum (PhABS) for their ability to promote MPC production under selective culture conditions. As the majority of "contaminating" cells in MPC cultures were represented by MSC-like cells, we hypothesized a role by differentiating agents present in the sera. Therefore, we tested a number of growth factors (hGF) and found that higher concentrations of FGF-2, EGF, PDGF-AB, and VEGF-A as well as lower concentration of IGF-1 give sub-optimal MPC recovery. Gene expression analysis of hGF receptors was also carried out both in MSCs and MPCs, suggesting that FGF-2, EGF, and PDGF-AB could act promoting MSC proliferation, while VEGF-A contribute to MSC-like cell contamination, triggering MPC differentiation. Here we demonstrated that managing hGF contents, together with applying specific receptors inhibitors (Erlotinib-HCl andNintedanib), could significantly mitigate the batch-to-batch variability related to serum supplementation. These data represent a fundamental milestone in view of manufacturing MPC-based medicinal products.
authors: Department of Clinical and Experimental Medicine, University of Pisa Pisa, Italy.
authors: Marina
authors: M
authors: Montali
authors: Department of Clinical and Experimental Medicine, University of Pisa Pisa, Italy.
authors: Serena
authors: S
authors: Barachini
authors: Department of Clinical and Experimental Medicine, University of Pisa Pisa, Italy.
authors: Francesca M
authors: FM
authors: Panvini
authors: Department of Surgical, Medical and Molecular Pathology and Critical Care Medicine, University of Pisa Pisa, Italy.
authors: Vittoria
authors: V
authors: Carnicelli
authors: Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa Pisa, Italy.
authors: Franca
authors: F
authors: Fulceri
authors: Department of Surgical, Medical and Molecular Pathology and Critical Care Medicine, University of Pisa Pisa, Italy.
authors: Iacopo
authors: I
authors: Petrini
authors: Department of Clinical and Experimental Medicine, University of Pisa Pisa, Italy.
authors: Simone
authors: S
authors: Pacini
keywords: Erlotinib; Nintedanib; bone marrow culture; cell-based medicinal product; growth factors; human serum; mesangiogenic progenitor cells; mesenchymal stromal cells