Culturing Human Neural Stem Cells and Quantifying Lentiviruses to Study Autism.
Milagros Mulero1, Leana Ramos1, Vadym Trokhymchuk1, Deliabell Hernandez1, Alexis Tapanes-Castillo1, Derek Dyxkhoorn2
1School of Science, Technology, and Engineering Management, St. Thomas University, Miami Gardens, FL 33054; 2Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, FL 33136
Autism spectrum disorder (ASD) is partly caused by problems in neuronal communication. Differences in the RNA expression levels of the synaptic protein Discs large homolog-associated protein 1 (DLGAP1) have recently been associated with ASD. The long term goal of this project is to analyze how human neurons react to reduced DLGAP1 antisense RNA levels. To accomplish this, we cultured neural progenitor cells (NPCs) from autistic and control patients. These neural progenitor cells, which were derived from induced pluripotent stem cells that originated from patient skin biopsies, were differentiated towards a glutamatergic fate. In separate experiments, done at the University of Miami, six lentiviruses designed to degrade DLGAP1 antisense RNA transcripts were produced. By generating a standard curve, we quantified the number of virus particles present in each sample. In the future, autistic and control neural progenitor cells will be infected with lentiviruses to analyze how reducing DLGAP1 antisense RNA levels affects autistic cells.