MePR: A Novel Human Mesenchymal Progenitor Model with Characteristics of Pluripotency

Abstract

Human embryo stem cells or adult tissues are excellent models for discovery and characterization of differentiation processes. The aims of regenerative medicine are to define the molecular and physiological mechanisms that govern stem cells and differentiation. Human mesenchymal stem cells (hMSCs) are multipotent adult stem cells that are able to differentiate into a variety of cell types under controlled conditions both in vivo and in vitro, and they have the remarkable ability of self-renewal. hMSCs derived from amniotic fluid and characterized by the expression of Oct-4 and Nanog, typical markers of pluripotent cells, represent an excellent model for studies on stemness. Unfortunately, the limited amount of cells available from each donation and, above all, the limited number of replications do not allow for detailed studies. Here, we report on the immortalization and characterization of novel mesenchymal progenitor (MePR) cell lines from amniotic fluid-derived hMSCs, whose biological properties are similar to primary amniocytes. Our data indicate that MePR cells display the multipotency potential and differentiation rates of hMSCs, thus representing a useful model to study both mechanisms of differentiation and pharmacological approaches to induce selective differentiation. In particular, MePR-2B cells, which carry a bona fide normal karyotype, might be used in basic stem cell research, leading to the development of new approaches for stem cell therapy and tissue engineering.

Secretome profiling of cytokines and growth factors reveals that neuro-glial differentiation is associated with the down-regulation of Chemokine Ligand 2 (MCP-1/CCL2) in amniotic fluid derived-mesenchymal progenitor cells

“…1A). The acquisition of the typical neural phenotype, previously described by Miceli et al [24], was further confirmed by the increase, upon MePR-2B differentiation, of intracellular calcium, a typical hallmark of neuronal differentiation (Fig. 1B).…”

“…MePR-2B cells can be cultivated as undifferentiated cells but, upon appropriate stimuli, can undergo myogenic, adipogenic, osteogenic, chondrogenic and neuro-glial differentiation [24]. In this work, a targeted secretome characterization was performed on undifferentiated cells (MePR-2B) and after the acquisition of morphological and physiological features typical of neuro-glial differentiation (MePR-2B/D) [24]. As expected, a typical spindle-shaped, fibroblastic-like morphology was observed for undifferentiated MePR-2B cells while, following differentiation, a neurallike morphology was acquired as attested by the appearance of a translucid shape with long extensions (Fig.…”

Identification and characterization of PKF118-310 as a KDM4A inhibitor

“…Human colon cancer HCT-116 (ATCC, VA, USA) and HCT-116 p53 ¡/¡ (DSMZ, Braunschweig, Germany) cells were propagated in McCoy's 5A medium (Euroclone, Milan, Italy) with 10% fetal bovine serum (FBS) (Euroclone), 2 mM L-glutamine (Euroclone) and antibiotics (100 U/ml penicillin, 100 lg/ml streptomycin) (Euroclone). 38 Human leukemic monocyte lymphoma U937 cells 39 (ATCC) and mesenchymal progenitor (MePR2B) cells 40 were propagated in RPMI 1640 medium containing 4.5 g/L glucose (Euroclone) supplemented with 10% FBS (Euroclone), 100 U/mL penicillin-streptomycin (Euroclone) and 2 mM L-glutamine (Euroclone). Cells were stimulated with drugs for 12 h, 24 h, 48 h, and 24 h C 24 h (drugs were added again 24 h post-24 h stimulation).…”

SIRT1 Inhibition Affects Angiogenic Properties of Human MSCs

“…Cells were first grown for 24 h and then infected with the rLV.H1.sh2Sirt1.EF1.GFP Lentivirus, with a 2.5 MOI, overnight as described by Miceli et al [27]. …”

“…Cell cycle distribution was assessed with a FACScalibur flow cytometer (Becton Dickinson, San Jose, CA, USA), and 10.000 cells were analyzed by ModFit version 3 Technology (Verity Software House, Topsham, ME, USA) and Cell Quest (Becton Dickinson, San Jose, CA, USA) [27]. …”