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Objective:
To evaluate the ability to isolate lung progenitor cells from amniotic fluid through manual isolation and c-kit targeted magnetic sorting. The secondary objective evaluated amniotic fluid stem cell (AFSC) growth in an engineered biomimetic lung scaffold.
Design:
AFSCs, obtained during third trimester amniocentesis, were isolated by two mechanisms: identification with manual removal and c-Kit (CD117) magnetic sorting. These cells were cultured and analyzed by immunofluorescence and fluorescent activated cell sorting (FACS) for pluripotency, mesenchymal, and hematopoietic markers. AFSCs were seeded into a decellularized scaffold and cultured in a bioreactor for 72 hours. Apoptosis and proliferation assays were performed.
Setting:
Laboratory
Patients:
N/A
Interventions:
N/A
Main Outcome Measure:
The isolation of relevant AFSCs expressing lung progenitor markers with the ability to seed a biomimetic scaffold.
Results:
FACS for manually isolated cells exhibited pluripotent and mesenchymal markers, but lacked hematopoietic markers. FACS for magnetic sorting based on CD117 demonstrated a similar profile. Immunofluorescence samples confirmed the FACS results. The manually isolated AFSCs seeded in the biomimetic lung scaffold were viable and proliferating after 72 hours in the perfusion bioreactor.
Conclusions:
CD117 magnetic sorting and mechanical isolation techniques yielded cells that expressed pluripotent and mesenchymal stem cell markers, while lacking hematopoietic markers. The AFSCs within the decellularized lung scaffold were maintained and proliferated while in the bioreactor. In conclusion, AFSCs have a high potential to be clinically translatable because of their patient specificity and accessibility early in pregnancy. However, further experimentation is necessary.