Isolation and characterization of mesenchymal cells from human fetal membranes

Abstract

Bone marrow (BM) multipotent mesenchymal stromal cells (MSCs) present with multipotent differentiation potential and immunomodulatory properties. As an alternative to bone marrow, we have examined fetal membranes, amnion and chorion, of term human placenta as a potential source of multipotent MSCs. Here we show that amnion mesenchymal cells (AMCs) and chorion mesenchymal cells (CMCs), isolated by mechanical separation and subsequent enzymatic digestion, demonstrate plastic adherence and fibroblast-like morphology and are able to form colonies that could be expanded for at least 15 passages. By FACS analysis, AMCs and CMCs were shown to be phenotypically similar to BM-MSCs and, when cultured in differentiation media, they demonstrated high morphogenetic plasticity by differentiating into osteocytes, chondrocytes and adipocytes. In an attempt to isolate cells with MSC characteristics from human fetal membranes, AMCs and CMCs expressing CD271 were enriched by immunomagnetic isolation and were demonstrated to possess higher clonogenic and osteogenic differentiation potential than CD271-depleted fractions. Based on these findings, amnion and chorion can be considered as a novel and convenient source of adult MSCs.

Placental Mesenchymal Stromal Cells Derived from Blood Vessels or Avascular Tissues: What Is the Better Choice to Support Endothelial Cell Function?

“…Both cell types lack expression of MSCA-1 and CD271, which have been suggested as markers for an especially clonogenic subpopulation of bone marrow MSCs [24][25][26][27]. While MSCA-1 and AP are known to be only expressed by bone-marrow-derived MSCs [24], the expression of CD271 has been described in a small subpopulation of av-MSCs before [17].…”

“…av-MSCs were isolated from the placental amnion as described earlier [14,17]. Amnion and chorion were manually separated and washed with 0.9% saline (Fresenius Kabi) supplemented with 150 IU/mL penicillin, 150 mg/mL streptomycin (both from PAA Laboratories), and 0.4 mg/mL amphotericin B (Gibco, Invitrogen).…”

In Vitro Studies of Horse Umbilical Cord Matrix-Derived Cells: From Characterization to Labeling for Magnetic Resonance Imaging

“…Chondrogenic differentiation was assessed in monolayer culture by incubating cells for 3 weeks in Soncini et al (2007) modified medium [19], composed of DMEM lowglucose containing 100 nM dexamethasone, 50 µg/ml Lascorbic acid 2-phosphate, 1 mM sodium pyruvate (BDH Chemicals Ltd., Poole, UK), 40 µg/ml proline, ITS (5 µg/ml Insulin, 5 µg/ml Transferrin, 5 ng/ml Sodium Selenite; Sigma) and 5 ng/ml TGF-β3 (Peprovet, DBA, Italia). Noninduced control cells were cultured for the same time in standard control medium.…”

Potential of Human Fetal Chorionic Stem Cells for the Treatment of Osteogenesis Imperfecta

“…For example, it is essential to have a source of stem cells that have high therapeutic potential and are easily accessible for clinical use. Extra-embryonic fetal tissues, such as the placenta, are readily available either from termination of pregnancy or surplus tissue at routine prenatal diagnostic procedures [17,18], or at term delivery [19,[20][21][22]. Recently, we have shown human fetal early chorionic stem cells (e-CSC) isolated from human placental tissue accelerated tissue repair in dermal excision skin wounds and improved bone quality and plasticity in oim mice.…”