Instead, fetal CV-MSCs require isolation under specific conditions, which has implications for clinical trials using placental MSC. We conclude that in standard media, fetal chorionic villi-derived MSC (CV-MSC) do not grow readily, whereas maternal MSC proliferate to result in maternal overgrowth during culture. Comparison of MSC populations within the same placenta confirmed fetal to be smaller, more osteogenic and proliferative than maternal MSC. To isolate pure fetal cells, we validated a novel isolation procedure comprising focal dissection from the cotyledonary core, collagenase/dispase digestion and explant culture in endothelial growth media that selected, and provided a proliferative environment, for fetal MSC.
Cultures in standard media rapidly became composed entirely of maternal cells despite isolation from fetal villi. We first demonstrated reproducible maternal cell contamination in MSC cultures from all chorionic anatomical locations tested. Fetal and maternal cell identities were quantitated in gender-discordant pregnancies by XY chromosome fluorescence in situ hybridization. MSCs were isolated via a range of methods in combination selection from various chorionic regions, different commercial media, mononuclear cell digest and/or explant culture. We determined isolation conditions to yield fetal and separately maternal MSC during ex vivo expansion from human term placenta. Our recent systematic review concluded that only a small minority of placental MSC publications report fetal/maternal origin, and failed to discern a specific methodology for isolation of fetal MSC from term villi. Cultured placental MSCs are confounded by a high frequency of maternal cell contamination.
Human placenta is rich in mesenchymal stem/stromal cells (MSC), with their origin widely presumed fetal.
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