Isolation of Lineage-Restricted Neuronal Precursors from Multipotent Neuroepithelial Stem Cells

Abstract

We have identified a neuronal-restricted precursor (NRP) cell that expresses E-NCAM (high polysialic-acid NCAM) and is morphologically distinct from multipotent neuroepithelial (NEP) cells (Kalyani et al., 1997) and spinal glial progenitors (Rao and Mayer-Proschel, 1997). NRP cells self renew over multiple passages in the presence of fibroblast growth factor (FGF) and neurotrophin-3 (NT-3) and differentiate in the presence of retinoic acid and the absence of FGF into postmitotic neurons. NRP cells can also be generated from multipotent E10.5 NEP cells. Clonal analysis shows that NRP cells arise from a NEP progenitor that generates other restricted CNS precursors. The NEP-derived NRPs undergo self renewal and can differentiate into multiple neuronal phenotypes. Thus, a direct lineal relationship exists between multipotential NEP cells and more restricted neuronal precursor cells present in vivo at E13.5 in the spinal cord.

SDF1α/CXCR4 Signaling, via ERKs and the Transcription Factor Egr1, Induces Expression of a 67-kDa Form of Glutamic Acid Decarboxylase in Embryonic Hippocampal Neurons

“…Brains were cryoprotected by immersion first in 20% sucrose for 24 h and then in 30% sucrose for 24 h. The samples were embedded in Tissue-Tek (Sakura, Torrance, CA), and cryostat sections were cut in the coronal plane at a thickness of 7 m. For immunohistochemical staining, the sections were first incubated with blocking solution (5% normal goat serum and 0.1% Triton X-100 in PBS, pH 7.4) for 1 h and then with anti-GAD67 (1:200; Chemicon) in blocking solution overnight at room temperature. After washing with PBST (0.1% Triton X-100 in PBS, pH 7.4), the sections were incubated with Alexa Fluor488-conjugated secondary antibodies (1:200; Invitrogen) at room temperature for 1 h. Staining procedures for cells were similar to those described previously (32). Briefly, the cells were fixed using 3% paraformaldehyde plus 1% glutaraldehyde in PBS for 30 min.…”

Cell Therapy for Multiple Sclerosis

“…The notion that glial committed neural stem cells may combine the capacity for self-renewal and plasticity of stem cells, together with the remyelinating properties of OPCs, led to identification of such precursors in the developing brain [134]. Expression of PSA-NCAM on the cell membrane has been associated with stem cell commitment to neuronal or glial fate, depending on time and place in development [135][136][137]. Indeed, such PSA-NCAM+ glial precursors, growing as neurospheres and also termed as oligospheres [135,138], efficiently myelinated the brains of shi mice [138][139][140], remyelinated 95 to 100% of axons following local injection into the dorsal columns of rats [141], and efficiently migrated along inflamed white matter tracts of rats with EAE [94,95].…”

Whole Genome Analysis of Human Neural Stem Cells Derived from Embryonic Stem Cells and Stem and Progenitor Cells Isolated from Fetal Tissue

“…To our knowledge, no direct comparisons between these populations in humans have been made. Several additional multipotent populations and dividing progenitors with a more restricted phenotype have also been identified in both mouse and human tissue [23][24][25]. Goldman and colleagues, for example, have isolated a glial progenitor population that has the ability to differentiate into oligodendrocytes and astrocytes when transplanted into a shiverer mouse model [26].…”