LHX6 On GABAergic Interneurons Fate Determination In Xeno-free System

As major inhibitory neurons in cerebral cortex,gamma-aminobutyric acid(GABAergic)intemeurons contributes to central never sysytems.Previous studies suggested that the transcriptional factor Lhx6 regulated the subtypes and migration’ of GABAergic intemeurons.Although breakthoughs have been made on the mechanism of the regulation of Lhx6,all of the researches were based on animal models.Besides,the regulation mechanism of human gene still remains unknown.Hipsc(Human induced pluripotent stem cells)has been reported having the potential to differentiate to all kinds of cells.To date,there have been plenty protocols involving in neuron differentiation from hipsc.However,much of the present protocols established in animal feeders,which introduced xeno factors and is not suit in human research and future clinical application.In our research,we construct an optimized xeno-free system,which aimed at directly differentiating hipsc to forebrain GABAergio intemeuons.Compared 5 different culture systems,we screened the most efficient culturing system.Besides,the only addition in patterning is a compound,Purmorphamine.The highly purified GABAergic intemeurons were yielded under our optimized xeno-free system with the addition of purmorphamine,a SHH substitution.Under xeno-free condition,we established inducible Lhx6 0overexpress hips cell line.Inducible Lhx6 overexpress hips cell line was then directly differentiate to GABAergic intemeurons.And the migration and subtype percentage were analyzed by brain slice co-culture and in vivo transplantation.Combine all these results,we found that Lhx6 could improve the percentage of PV and SST from GABA interneurons and ventralized the neurons.Our study makes great efforts to explore cerebral cortex development and search the therapy for neurodegenerative diseases.