Human fetal cortical and striatal neural stem cells generate region‐specific neurons in vitro and differentiate extensively to neurons after intrastriatal transplantation in neonatal rats

Abstract

Human fetal brain is a potential source of neural stem cells (NSCs) for cell replacement therapy in neurodegenerative diseases. We explored whether NSCs isolated from cortex and striatum of human fetuses, aged 6-9 weeks post-conception, maintain their regional identity and differentiate into specific neuron types in culture and after intrastriatal transplantation in neonatal rats. We observed no differences between cortex- and striatum-derived NSCs expanded as neurospheres in proliferative capacity, growth rate, secondary sphere formation, and expression of neural markers. After 4 weeks of differentiation in vitro, cortical and striatal NSCs gave rise to similar numbers of GABAergic and VMAT2- and parvalbumin-containing neurons. However, whereas cortical NSCs produced higher number of glutamatergic and tyrosine hydroxylase- and calretinin-positive neurons, several-fold more neurons expressing the striatal projection neuron marker, DARPP-32, were observed in cultures of striatal NSCs. Human cortical and striatal NSCs survived and migrated equally well after transplantation. The two NSC types also generated similar numbers of mature NeuN-positive neurons, which were several-fold higher at 4 months as compared to at 1 month after grafting. At 4 months, the grafts contained cells with morphologic characteristics of neurons, astrocytes, and oligodendrocytes. Many of neurons were expressing parvalbumin. Our data show that NSCs derived from human fetal cortex and striatum exhibit region-specific differentiation in vitro, and survive, migrate, and form mature neurons to the same extent after intrastriatal transplantation in newborn rats.

Human midbrain precursors activate the expected developmental genetic program and differentiate long-term to functional A9 dopamine neurons in vitro. Enhancement by Bcl-XL

“…2), matching what has been described for FB-derived hNSl cells [42,72], and human cortical, striatal and VM neurosphere cultures [26,63]. Nestin + cells only disappeared completely by d30, enforcing the view that hNSCs require long differentiation times in vitro to mature (present work and see also [26,40,63]). The forced expression of Bcl-X L did not alter the down-regulation of these markers (Supplementary information Fig.…”

“…Patterning preservation has been demonstrated in cortical, striatal and midbrain neurospheres [57,58,63], but there is only indirect evidence for FB or VM v-myc hNSC lines [34][35][36][40][41][42]64], and the cell lines have never been studied head-to-head. In the present work we first clarified that long-term cultured FB and VM hNSCs differentially express early neuroectodermal markers (PAX6, EN1), and generate neurons showing differential expression of A9-DAn genes (such as GIRK2) (Fig.…”

Schwann-cell cylinders grown inside hyaluronic-acid tubular scaffolds with gradient porosity

“…However, in vivo experiments with glia, neuronal precursors and stem cells [29,30] show a significant loss of transplanted cells at the site of grafting, leading to a poor restoration of functional properties [31]. In order to face this problem, porous scaffolds with tailored properties and structure can be used to provide to the cells a more friendly environment and to prevent them from spreading out of the lesion site [18,19,32].…”

Adult human neural stem cell therapeutics: Current developmental status and prospect

“…In particular, human ESCderived NSCs, injected into the ischemic penumbra region in rat brains with ischemic stroke, have been reported to infarction area, and showed recovery of motor function [18] . When human fetal NSCs were transplanted into ischemic lesions of rodent brains, they migrated toward the injured regions and differentiated into neurons [63,64] . move to the lesions, and improve motor performances [62] .…”