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

Abstract

Studying how the fetal spinal cord regenerates in an ex vivo model of spina bifida repair may provide insights into the development of new tissue engineering treatment strategies to better optimize neurologic function in affected patients. Here, we developed hydrogel surgical patches designed for prenatal repair of myelomeningocele defects and demonstrated viability of both human and rat neural progenitor donor cells within this three-dimensional scaffold microenvironment. We then established an organotypic slice culture model using transverse lumbar spinal cord slices harvested from retinoic acid–exposed fetal rats to study the effect of fibrin hydrogel patches ex vivo. Based on histology, immunohistochemistry, gene expression, and enzyme-linked immunoabsorbent assays, these experiments demonstrate the biocompatibility of fibrin hydrogel patches on the fetal spinal cord and suggest this organotypic slice culture system as a useful platform for evaluating mechanisms of damage and repair in children with neural tube defects.

Amniotic fluid‐derived stem cell potential for therapeutic and surgical use: A review of the literature

“…More recently, induction of pluripotent stem cells into keratinocytes, for dorsal skin coverage of spina bifida in ex vivo models has succeeded 38 . Production of injectable hydrogel patches for prenatal spina bifida repair has shown biocompatibility when human AFSCs are induced to form neural progenitor cells 39 . These cells were found to maintain viability within a 3D environment—a major step forward.…”

“…However, deeper investigation into the effectiveness of AFSCs derived from healthy fetuses verses those with neural tube defects, specifically, yet remains. Success for AFSCs in treating ex vivo spina bifida models with hydrogel patches, 39 provides hope for future surgical AFSC use. “Trans‐amniotic stem cell therapy” or TRASCET, has been of recent interest regarding the use of AFSCs in treating spina bifida.…”

“…38 Production of injectable hydrogel patches for prenatal spina bifida repair has shown biocompatibility when human AFSCs are induced to form neural progenitor cells. 39 These cells were found to maintain viability within a 3D environment-a major step forward.…”

Progress in application of natural scaffold materials in pulp regeneration

“…Because it is derived from autologous, fibrin itself or its degradation products are not toxic or produce inflammatory response. Fibrin is a natural scaffold material derived from animal proteins, which has good mechanical and physicochemical properties and can be used as an effective carrier for cells and signaling molecules, providing an environment for cell adhesion in the form of hydrogels, and regulating the differentiation of cell growth [39][40][41] . Nisarat et al [42] injected fibrin hydrogel into an endodontically devoid root canal, which affected cell morphology and differentiation function in contact with dentin compared to the empty root canal, and promoted tissue growth into the root canal, with newly formed tissue similar to normal pulp.…”

Human Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Neural Differentiation of Neural Progenitor Cells

“…First, to investigate the differentiation behavior of multipotent ADMSCs in neurogenesis media, which are the conditions required for EV isolation, immunocytochemistry (ICC) analysis was conducted for staining of neuronal cell-specific proteins (Figure 2A). Differentiated ADMSCs were visualized with Nestin as a neural progenitor cell marker, glial fibrillary acidic protein (GFAP) (an intermediate filament) as an astrocyte marker, and microtubule-associated protein 2 (MAP2) as a mature neuron marker [24][25][26][27][28][29]. First, to investigate the differentiation behavior of multipotent ADMSCs in neurogenesis media, which are the conditions required for EV isolation, immunocytochemistry (ICC) analysis was conducted for staining of neuronal cell-specific proteins (Figure 2A).…”