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Background: Donor neural stem cells (NSCs) can lead to fetal spinal cord repair, in an animal model of spina bifida. We sought to determine whether NSCs could be isolated from the amniotic fluid in the setting of neural tube defects (NTDs), as a preclinical requirement for autologous cell sourcing.
Methods: After IACUC approval, 62 pregnant Sprague-Dawley dams were divided into experimental (n=42) and control (n=20) groups, depending on prenatal exposure to retinoic acid for the induction of fetal NTDs. All animals were killed before term for analysis (n=685 fetuses). Amniotic fluid samples from both groups underwent epigenetic selection for NSC, followed by exposure to neural differentiation media. Representative cell samples underwent multiple morphological and phenotypical analyses at different time points.
Results: None of the 267 control fetuses had a structural abnormality, whereas at least one NTD was present in 52% (217/418) of the experimental fetuses, namely either an isolated spina bifida (n=144), an isolated encephalocele (n=24), or a combination of the two (n=49). Only amniotic samples from fetuses with a NTD yielded cells with typical neural progenitor phenotype. These cells responded to differentiation media displaying elaborate morphological changes, along with expression of Glial Fibrillary Acidic Protein (GFAP), Beta-Tubulin, and/or O4, concomitantly with Nestin downregulation, pointing to differentiation into different neural lineages.
Conclusion: The amniotic fluid can harbor neural progenitor cells in the setting of experimental neural tube defects. The amniotic fluid may be a practical source of autologous neural progenitor cells applicable to novel forms of therapy for spina bifida.