| Autor: |
Gonzalez H; Department of Internal Medicine, Division of Infectious Diseases, Rush University Medical Center, Chicago, IL 60612, USA.; Department of Microbial Pathogens and Immunity, Rush Medical College, Chicago, IL 60612, USA., Narasipura SD; Department of Microbial Pathogens and Immunity, Rush Medical College, Chicago, IL 60612, USA., Shull T; Department of Microbial Pathogens and Immunity, Rush Medical College, Chicago, IL 60612, USA.; Division of Epidemiology and Biostatistics, School of Public Health, University of Illinois, Chicago, IL 60608, USA., Shetty A; Illinois Mathematics and Science Academy, Aurora, IL 60506, USA., Teppen TL; Molecular Neurobiology Division, Rush Alzheimer's Disease Center, Rush University, Chicago, IL 60612, USA., Naqib A; Genome Core Facility, Rush University, Chicago, IL 60612, USA., Al-Harthi L; Department of Microbial Pathogens and Immunity, Rush Medical College, Chicago, IL 60612, USA. |
| Abstrakt: |
Human inducible pluripotent stem cell (hiPSC)-derived astrocytes (iAs) are critical to study astrocytes in health and disease. They provide several advantages over human fetal astrocytes in research, which include consistency, availability, disease modeling, customization, and ethical considerations. The generation of iAs is hampered by the requirement of Matrigel matrix coating for survival and proliferation. We provide a protocol demonstrating that human iAs cultured in the absence of Matrigel are viable and proliferative. Further, through a side-by-side comparison of cultures with and without Matrigel, we show significant similarities in astrocyte-specific profiling, including morphology (shape and structure), phenotype (cell-specific markers), genotype (transcriptional expression), metabolic (respiration), and functional aspects (glutamate uptake and cytokine response). In addition, we report that, unlike other CNS cell types, such as neuronal progenitor cells and neurons, iAs can withstand the absence of Matrigel coating. Our study demonstrates that Matrigel is dispensable for the culture of human iPSC-derived astrocytes, facilitating an easy, streamlined, and cost-effective method of generating these cells. |
