Neural progenitor cells derived from the adult rat subventricular zone: characterization and transplantation.
In order to fully characterize and determine the therapeutic potential of adult neural progenitor cells (NPCs), it is important to be able to isolate and study NPCs from animals such as rats, in which there are existing models of brain injury and disease. The focus of this study was to characterize the cultivation, differentiation, and transplantation of adult rat NPCs isolated from the subventricular zone of the lateral ventricles. We examined strategies for cell purification using a Percoll density gradient, and cell expansion using a range of maintenance medium and plating densities. Purification by Percoll gradient enriched a population of cells expressing nestin and SOX2, but resulted in a significant reduction in neurosphere generation. Culturing adult rat NPCs in Neurobasal-A media and plating at 200,000 cell/ml resulted in a higher percentage of cells surviving to generate neurospheres compared to culture in DMEM/F12 or NS-A media. On induction of differentiation, adult rat NPCs were capable of generating neurons, astrocytes, and oligodendrocytes in vitro that survived for up to 8 weeks, demonstrating multipotentiality of these cells. In addition, a population of cells continued to proliferate during the initial phase of differentiation, suggesting the presence of two populations of NPCs during differentiation. Cultured adult rat NPCs also survived and differentiated into astrocytes 6 weeks after transplantation into the striatum of the normal adult rat brain. In conclusion, we have optimized techniques that allow for the routine isolation, culture, and transplantation of multipotent NPCs derived from the adult rat SVZ.
Chen
Kevin
K
Department of Pharmacology and Clinical Pharmacology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
Hughes
Stephanie M
SM
Connor
Bronwen
B
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000032:analysis; D024002:HMGB Proteins / Q000032:analysis; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000032:analysis; D020547:Lateral Ventricles / Q000166:cytology*; D008297:Male; D009419:Nerve Tissue Proteins / Q000032:analysis; D064231:Nestin; D009474:Neurons / Q000737:chemistry / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D055748:SOXB1 Transcription Factors; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology*; D014157:Transcription Factors / Q000032:analysis
D004268:DNA-Binding Proteins; D024002:HMGB Proteins; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D055748:SOXB1 Transcription Factors; C527784:Sox2 protein, rat; D014157:Transcription Factors