In vitro and In vivo analysis of gene expression changes in mouse neural progenitor cells derived from embryonic stem cells after exposure to specific molecules or grafting into traumatic spinal cord injury

Comparison of genomic data from neural progenitor cells derived from mouse embryonic stem cells under different experimetnal conditions in vitro and invivo. We conducted genome-wide RNA sequencing of immunoprecipitated specific ribosome-associated mRNA using RiboTag methods from: (i) mouse embryonic stem cell (ESC), (ii) derived neural progenitor cells, (iii) differentiated neural progenitor cells (in vitro), (iv) grafted neural progenitor cells (recovered from different in vivo tissue enivornments - healthy spinal cord, spinal cord injury lesions) and (v) host astrocytes using GFAp-Cre RiboTag mice. Mouse ESC were derived from the inner cell mass of E3.5 blastocyst stage embryos generated from crosses of male homozygous B6N.129-Rpl22tm1.1 Psam/J “Ribotag” mice to females hemizygous for a dominant, maternal effect cre allele, B6.Cg-Tg(SOX2-cre)1Amc/J and heterozygous for the “RiboTag” allele. NPC were derived by neural induction and expansion protocols. Neural progenitors were differenetaitatd in vitro for 4 days by removal of progenitor maintaing mitogens (EGF/FGF) and treatment with either CNTF, IGF-1 or FBS in different concentrations. NPC mRNA was recoveved using the RiboTag protocol. NPC were grafted into young adult mice that underwent severe crush SCI at thoracic level T10. NPC grafting was performed at 2 days after injury. NPC mRNA was recoveved using the RiboTag protocol at 5 and 14 days after injury.