Purpose: This study was aimed at determining whether fetal tissue constructs can be engineered from cells normally found in the amniotic fluid. Methods: A subpopulation of morphologically distinct cells was isolated mechanically from the amniotic fluid of pregnant ewes (n = 5) and expanded selectively. Its lineage was determined by immunofluorescent staining against multiple intermediate filaments and surface antigens. Proliferation rates were determined by both oxidation and total DNA assays and compared with immunocytochemically identical adult and fetal sheep cells. Statistical analysis was by analysis of variance for repeated measures (ANOVA). After expansion, the amniocytes were seeded onto a polyglycolic acid polymer/poly-4-hydroxybutyrate scaffold. The resulting construct was analyzed by both optical and scanning electron microscopy. Results: The immunocytochemical profile of expanded amniocytes was consistent with a mesenchymal, fibroblast/myofibroblast cell lineage. These cells proliferated significantly faster than comparable fetal and adult cells in culture. Amniocyte construct analysis showed dense, confluent layers of cells firmly attached to the scaffold, with no evidence of cell death. Conclusions: (1) Subpopulations of fetal mesenchymal cells can be isolated consistently from the amniotic fluid. (2) Mesenchymal amniocytes proliferate more rapidly in vitro than comparable fetal and adult cells. (3) Mesenchymal amniocytes attach firmly to polyglycolic acid polymer. The amniotic fluid can be a reliable and practical source of cells for the engineering of select fetal tissue constructs.
Bibliographical noteFunding Information:
This research was supported by grants from the United States Surgical Corporation and The Children's Hospital Surgical Foundation.
- Amniotic fluid
- Birth defects
- Congenital anomalies
- Fetal surgery
- Tissue engineering