Existing treatments for myelomeningocele repair consist

Existing treatments for myelomeningocele repair consist of pre- or post-natal surgical closure of the defect. Prenatal intervention has been shown to be more effective than postnatal surgery because it can prevent or ameliorate sequels (Adzick et al., 2011; Adzick et al., 1998; Meuli et al., 1997; Meuli et al., 1995). Clearly, in utero repair of myelomeningocele is neither a complete remedy nor free of risk for mother and fetus (Adzick et al., 2011; Meuli and Moehrlen, 2014). Despite the benefits of surgical repair in preventing further injury, neurological outcomes remain a critical concern (Gebb et al., 2015) because of the damage incurred before closure of the defect. In spite of the low 3% perinatal mortality rate and possible benefits of in-utero repair of myelomeningocele, 36% of patients after surgery had mild to severe impairment (Danzer et al., 2010; Hisaba et al., 2012). Therefore, new therapeutic approaches concomitant to fetal surgery are needed to improve locomotive and cognitive functions in these babies. During the last 10years, researchers made great efforts toward the development of neural stem cell-based therapeutic approaches for spinal cord regeneration (Barnabé-Heider and Frisén, 2008; Li and Lepski, 2013; Mothe and Tator, 2013; Parati et al., 2004; Sandner et al., 2012). Even with CSF\'s low cellularity (de Graaf et al., 2011), recent studies have reported that embryonic CSF contains neurogenic growth factors (Buddensiek et al., 2010; Zappaterra and Lehtinen, 2012). In pre-term babies with post-hemorrhagic hydrocephalus, CSF samples showed some circulating VCH-222 with neural progenitor properties (Krueger et al., 2006). Although technical difficulties exist in CSF sampling during gestation, the findings of neural progenitor cells (NPCs) in CSF strongly support its potential as an important source of stem cells for the new regenerative medicine-based therapeutic approaches for spinal cord injury repair. During prenatal surgical repair of myelomeningocele (Enriquez et al., 2012; Peiró et al., 2009a; Peiró et al., 2009b; Pellicer et al., 2007; Ruano et al., 2013a, 2013b), collection of fresh and well-preserved CSF samples from the cyst was feasible by gentle aspiration, thus permitting examination of its properties and cellular composition.
Materials and methods During the surgical repair of myelomeningocele in 6 fetuses between 20 and 26weeks of gestation, CSF samples were extracted from the cyst surrounding the exposed neural placode. The study was approved by the IRB Ethics Committees of CCHMC (IRB 2013-5324) and IRB of the Vall d\'Hebron University Hospital (protocol ID: PR(AMI)65/2013). Patients received detailed information about the experimental protocol and gave written consent for their participation.
Results By successful extraction of CSF samples from the cyst of the myelomeningocele during fetal surgery at mid gestation (Fig. 1), our protocol detected the presence of NPCs in CSF-derived primary culture by the expression of neural and pluripotent specific markers TBR2, SOX2, and CD15 using immunocytofluorescence, quantitative real-time PCR, and flow cytometry analysis.
Discussion
Conclusion The following are the supplementary data related to this article.
Funding source This study was supported by internal funds from Cincinnati Children\'s Hospital Medical Center, Pediatric General and Thoracic Surgery Division (SP15), and A. Bosch Foundation, Girona (Spain).
Acknowledgements
Introduction Pluripotent hESs have distinct chromatin structure that is homogeneous, decondensed, and enriched for active histone markers. Additionally, pluripotent hESs are in a transcriptionally globally hyperactive state, as they contain levels of total RNA and mRNA almost two times higher than differentiated neural progenitor cells (Efroni et al., 2008; Tee and Reinberg, 2014). Recent studies have shown that the splicing factors RBM9, ESRP1, SRSF2, MBNL1, MBNL2 and SON pay essential roles in regulating pluripotency by stimulating the precise splicing of specific variants or globally changing transcriptional expression (Cieply et al., 2016; Han et al., 2013; Lu et al., 2013; Venables et al., 2013; Ye and Blelloch, 2014). Depletion of the spliceosome-associated factor SON in hESs results in loss of pluripotency and cell death by the regulating the splicing of OCT4, PRDM14, E4F1 and MED24 (Lu et al., 2013). Therefore, the precise regulation of RNA splicing convincingly plays a pivotal role in driving pluripotency entry, maintenance, and exit.