Self-renewal and differentiation of reactive astrocyte-derived neural stem/progenitor cells isolated from the cortical peri-infarct area after stroke.
In response to stroke, subpopulations of cortical reactive astrocytes proliferate and express proteins commonly associated with neural stem/progenitor cells such as glial fibrillary acidic protein (GFAP) and Nestin. To examine the stem cell-related properties of cortical reactive astrocytes after injury, we generated GFAP-CreER(TM);tdRFP mice to permanently label reactive astrocytes. We isolated cells from the cortical peri-infarct area 3 d after stroke, and cultured them in neural stem cell medium containing epidermal growth factor and basic fibroblast growth factor. We observed tdRFP-positive neural spheres in culture, suggestive of tdRFP-positive reactive astrocyte-derived neural stem/progenitor cells (Rad-NSCs). Cultured Rad-NSCs self-renewed and differentiated into neurons, astrocytes, and oligodendrocytes. Pharmacological inhibition and conditional knock-out mouse studies showed that Presenilin 1 and Notch 1 controlled neural sphere formation by Rad-NSCs after stroke. To examine the self-renewal and differentiation potential of Rad-NSCs in vivo, Rad-NSCs were transplanted into embryonic, neonatal, and adult mouse brains. Transplanted Rad-NSCs were observed to persist in the subventricular zone and secondary Rad-NSCs were isolated from the host brain 28 d after transplantation. In contrast with neurogenic postnatal day 4 NSCs and adult NSCs from the subventricular zone, transplanted Rad-NSCs differentiated into astrocytes and oligodendrocytes, but not neurons, demonstrating that Rad-NSCs had restricted differentiation in vivo. Our results indicate that Rad-NSCs are unlikely to be suitable for neuronal replacement in the absence of genetic or epigenetic modification.
Department of Medicine, Stem Cell Core, University of Vermont, Colchester, Vermont 05446, USA.
Issei S
IS
Shimada
Matthew D
MD
LeComte
Jerrica C
JC
Granger
Noah J
NJ
Quinlan
Jeffrey L
JL
Spees
D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology; D001253:Astrocytes / Q000502:physiology*; D015153:Blotting, Western; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002452:Cell Count; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage; D002540:Cerebral Cortex / Q000166:cytology* / Q000502:physiology*; D002544:Cerebral Infarction / Q000473:pathology*; D004396:Coloring Agents; D005434:Flow Cytometry; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D020244:Infarction, Middle Cerebral Artery / Q000473:pathology; D008164:Luminescent Proteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D058953:Neural Stem Cells / Q000502:physiology* / Q000637:transplantation; D053764:Presenilin-1 / Q000037:antagonists & inhibitors / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D051881:Receptor, Notch1 / Q000037:antagonists & inhibitors / Q000235:genetics; D033581:Stem Cell Transplantation; D020521:Stroke / Q000473:pathology*; D000097573:Red Fluorescent Protein
D000963:Antimetabolites; D001973:Bromodeoxyuridine; D004396:Coloring Agents; D005904:Glial Fibrillary Acidic Protein; D008164:Luminescent Proteins; D053764:Presenilin-1; D051881:Receptor, Notch1; D000097573:Red Fluorescent Protein; C497762:Notch1 protein, mouse