Effect of Mother’s Age and Pathology on Functional Behavior of Amniotic Mesenchymal Stromal Cells—Hints for Bone Regeneration

Abstract

Human amnion-derived mesenchymal stromal cells (hAMSCs) are used increasingly in regenerative medicine applications, including dentistry. The aim of this study was to evaluate if hAMSCs from aged and pathological mothers could be affected in their phenotype and functional behavior. hAMSCs were isolated from placentas of women aged younger than 40 years (Group 1, n = 7), older than 40 years (Group 2, n = 6), and with pre-eclampsia (Group 3, n = 5). Cell yield and viability were assessed at isolation (p0). Cell proliferation was evaluated from p0 to p5. Passage 2 was used to determine the phenotype, the differentiation capacity, and the adhesion to machined and sandblasted titanium disks. hAMSCs recovered from Group 3 were fewer than in Group 1. Viability and doubling time were not different among the three groups. Percentages of CD29+ cells were significantly lower in Group 3, while percentages of CD73+ cells were significantly lower in Groups 2 and 3 as compared with Group 1. hAMSCs from Group 2 showed a significant lower differentiation capacity towards chondrogenic and osteogenic lineages. hAMSCs from Group 3 adhered less to titanium surfaces. In conclusion, pathology can affect hAMSCs in phenotype and functional behavior and may alter bone regeneration capacities.

The Oncogenic Theory of Preeclampsia: Is Amniotic Mesenchymal Stem Cells-Derived PLAC1 Involved?

“…The yield of hAMSCs from preeclamptic amniotic membranes (each of 5 cm 2 ) was comparable with that of non preeclamptic ones (preeclamptic: 6.51 ± 3.98 × 10 6 ; non-preeclamptic: 7.71 ± 5.33 × 10 6 [mean ± SD]). As indicated by the International Society for Cellular Therapy (ISCT), which proposed minimal criteria for identification of human MSCs [ 18 ], hAMSCs presented a fibroblast-like morphology in culture and were confirmed to be CD45 (−), CD11b (−), CD90 (+), CD73 (+), and CD105 (+) via flow cytometry [ 19 ].…”

“…hAMSCs were isolated from a piece of amniotic membrane of 5 cm 2 , and grown in advanced DMEM supplemented with 10% FBS, 55 μM β-mercaptoethanol, 1% L-glutamine, 1% penicillin/streptomycin, and 10 ng/mL epidermal growth factor (EGF) (Sigma-Aldrich, Milan, Italy), as previously described [ 19 , 41 ]. hAMSCs were subcultured until passage 2 and analyzed for their phenotype [ 19 ]. Passage 2 hAMSCs were seeded onto 60 mm Petri dishes at a cell density of 5 × 10 5 /dish.…”

Human Amniotic Mesenchymal Stem Cells and Fibroblasts Accelerate Wound Repair of Cystic Fibrosis Epithelium

“…They were subcultured until passage 5, when 80% of confluence was obtained. For phenotypic analysis, cells were then stained with fluorochrome-conjugated monoclonal antibodies against hematopoietic (CD14, CD34, CD45) or mesenchymal (CD29, CD73, CD105) markers, as previously published [ 41 ]. All antibodies were purchased from Invitrogen, Thermo Fisher Scientific (Waltham, MA, USA), and were conjugated with fluorescein isothiocyanate (FITC), except the antibody against CD73 that required an additive incubation with secondary antibody (FITC goat anti-mouse; Sigma-Aldrich, Milan, Italy) for 30 min at 4 °C.…”