Isolation, characterization, and differentiation of multipotent neural progenitor cells from human cerebrospinal fluid in fetal cystic myelomeningocele

Abstract

Despite benefits of prenatal in utero repair of myelomeningocele, a severe type of spina bifida aperta, many of these patients will still suffer mild to severe impairment. One potential source of stem cells for new regenerative medicine-based therapeutic approaches for spinal cord injury repair is neural progenitor cells (NPCs) in cerebrospinal fluid (CSF). To this aim, we extracted CSF from the cyst surrounding the exposed neural placode during the surgical repair of myelomeningocele in 6 fetuses (20 to 26weeks of gestation). In primary cultured CSF-derived cells, neurogenic properties were confirmed by in vitro differentiation into various neural lineage cell types, and NPC markers expression (TBR2, CD15, SOX2) were detected by immunofluorescence and RT-PCR analysis. Differentiation into three neural lineages was corroborated by arbitrary differentiation (depletion of growths factors) or explicit differentiation as neuronal, astrocyte, or oligodendrocyte cell types using specific induction mediums. Differentiated cells showed the specific expression of neural differentiation markers (βIII-tubulin, GFAP, CNPase, oligo-O1). In myelomeningocele patients, CSF-derived cells could become a potential source of NPCs with neurogenic capacity. Our findings support the development of innovative stem-cell-based therapeutics by autologous transplantation of CSF-derived NPCs in damaged spinal cords, such as myelomeningocele, thus promoting neural tissue regeneration in fetuses.

Hydrogel and neural progenitor cell delivery supports organotypic fetal spinal cord development in an ex vivo model of prenatal spina bifida repair

“…33,34 In addition, transdifferentiation of amniocytes into the neurogenic lineage and isolation of specific neurogenic cell populations that may be uniquely accessible in the MMC fetus may be viable alternatives for use in hydrogel patches. [35][36][37][38] Our rationale for selecting neural progenitors as the ideal cell type for the MMC patch was based on previous work showing that neural progenitors can improve axonal regeneration in both animal and slice culture models of adult spinal cord injury. [39][40][41][42] In addition to neural progenitor differentiation into neurons, the functional multipotency of these cells, including host cell integration, re-myelination of host neuronal circuitry, and constitutive secretion of neurotrophic and neuroprotective factors responsible for regrowth and axonal regeneration, has been demonstrated.…”

“… 33 , 34 In addition, transdifferentiation of amniocytes into the neurogenic lineage and isolation of specific neurogenic cell populations that may be uniquely accessible in the MMC fetus may be viable alternatives for use in hydrogel patches. 35 – 38 …”

Prenatal diagnosis, natural history, postnatal treatment and outcome of 222 cases of spina bifida: experience of a tertiary center

“…Fetal surgery is an important strategy to further improve long‐term outcome and to offer an alternative to termination of pregnancy in the most severe cases; however, it is associated with significant adverse maternal and fetal outcomes. Other new techniques, such as stem cell‐based therapeutic approaches for spinal cord regeneration and the use of cryopreserved human umbilical cord patch, need optimization. Our contemporaneous data on outcomes in conventionally managed SB cases provide a modern benchmark against which to compare such evolving strategies.…”

“…Other new techniques, such as stem cell-based therapeutic approaches for spinal cord regeneration 43,44 and the use of cryopreserved human umbilical cord patch, need optimization 45,46 . Our contemporaneous data on outcomes in conventionally managed SB cases provide a modern benchmark against which to compare such evolving strategies.…”

Language and Latino immigrants living with spina bifida: Social determinants of health – the missing dimension in quality of life research

“…Increasingly, investigational interventions are actively pursued in an effort to treat the issues associated with SB [18,19]. As innovative procedures are subsequently applied, their impact on QOL should be studied utilizing validated instruments.…”