Method of treatment
A stem cell and disease technology, applied in pharmaceutical formulations, medical raw materials derived from mammals, cells of the nervous system, etc., can solve problems such as no possibility of designing effective treatments
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Embodiment 1
[0106] Materials and methods
[0107] Mice. 14-3-3ζ carrying a gene trap construct containing a Geo reporter gene Gt(OST062)Lex and 14-3-3ζ Gt(OST390)Lex Mutant mice were obtained from Lexicon Genetics ES cell lines OST062 and OST390, respectively. 14-3-3ζ Gt(OST062)Lex The gene trap vector in mouse inserts the first intron of 14-3-3ζ, while 14-3-3ζ Gt(OST390)Lex Gene trap vector insertion in the second intron of 14-3-3ζ in mouse. ES cell lines were expanded and injected into SV129 blastocysts. The resulting germline transmission males were maintained on the SV129 background or backcrossed within 6 generations into C57 / Bl6 and BA1, BC backgrounds. qRT-PCR and western blots from whole tissue samples were employed to confirm complete KO of this gene in these mouse strains. The 14-3-3C genotype was determined by PCR amplification of genomic tail DNA using the primers detailed in Supplementary Table 1. The wild-type allele was amplified by 288bp (14-3-3ζ Gt(OST062)Lex ) or ...
Embodiment 2
[0139] Demonstration that NRP2-positive neuronal precursors generate hippocampal regions
[0140] To identify mature neurons derived from neuronal precursors and expressing Nrp1 or Nrp2, Nrp1 and Nrp2 line-traced mice were generated. To this end, Cre / RFP or Cre / GFP have been placed under Nrp1 or Nrp2 promoter expression ( Figure 16 ). Studies using these mice (from n=2 experiments, Figure 17 ) show for the first time that hippocampal neurons are derived from Nrp2-expressing neural stem cells.
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