Isolation of amniotic fluid-derived mesenchymal stem cells.

Abstract

“ARTEMISIA” Fetal-Maternal Medical Centre, Department Prenatal Diagnosis, Rome, ItalyReprint requests to: Alvaro Mesoraca“ARTEMISIA” Fetal-Maternal Medical CentreDepartment of Genetics and Molecular Biology, Viale Liegi, 45 - 00198 Rome - ItalyTel.: [phone]E-mail: [e-mail]l stem cells (MSC) are capable of differen-tiating into different mesenchymal lineages, includingadipose and connective tissue, bone and cartilage. Themost common source of MSCs for clinical use is humanadult bone marrow. Because the frequency and differen-tiating capacity of MSCs are decreasing with age, differ-ent fetal tissues have been studied as an alternativesource for MSCs. Recently, in ‘t Anker et al. (2003) andTsai et al. (2004) reported that second-trimester amniot-ic fluid (AF) is a novel and rich source of fetal MSCs use-ful for clinical applications. AF contains a heterogeneouspopulation of cells from fetal origin. The fetal MSCs inAF are probably deriving from the amnion membrane orother embryonic and extra-embryonic tissues during theprocess of fetal development and growth (in ‘t Anker etal., 2004). Aim of the studyThe purpose of this study was to investigate whether wewere able to isolate and cultivate MSC from fresh sec-ond-trimester amniotic fluid samples. Materials and methodsAmniotic fluid was collected from 8 second-trimesterpregnancies (mean gestational age, 16 weeks [range16-18 weeks]). Four mL of the AF samples were cen-trifugated for 10 minutes at 300g. Pellets were resus-pended and cultured in DMEM with low glucose supple-mented with 15% Fetal bovine serum and antibiotics.Culture flasks were incubated at 37°C, 5% CO