Long-term expansion of human neural progenitor cells by epigenetic stimulation in vitro.
Human neural progenitor cells (hNPCs) are currently believed to have important potential for clinical application and basic neuroscience research. In the present study, we have developed a new technique for expansion of human neural progenitor cells in vitro. We showed that the cultures of hNPCs in monolayer could keep the same features with that growing in neurospheres. These cells expressed the typical protein of neural progenitors, nestin, and could form neurons and astrocytes upon differentiation. Using this method, we achieved an exponential increase in cells number over a period of 240 days in vitro. We also confirmed these cells expressed the orphan nuclear receptor-related factor 1 (Nurr1). Furthermore, we acquired the GFP-expressing human neural progenitor cells using retroviral-mediated transgenic system. The results of present study indicate the feasibility of long-term in vitro expansion of human neural progenitor cells using the monolayer culture technique, which may be of value as vehicles for ex vivo gene transfer to the CNS and as a potential source for basic research of dopaminergic (DA) neurons development.
Zhang
Haiyan
H
Beijing Institute for Neuroscience, The Beijing Center of Neural Regeneration and Repairing, Capital University of Medical Sciences, Beijing 100054, China.
Zhao
Yongmei
Y
Zhao
Chunli
C
Yu
Shuang
S
Duan
Deyi
D
Xu
Qunyuan
Q
D015153:Blotting, Western / Q000379:methods; D001921:Brain / Q000166:cytology* / Q000196:embryology; D001973:Bromodeoxyuridine / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D044127:Epigenesis, Genetic / Q000502:physiology*; D005333:Fetus; D005434:Flow Cytometry / Q000379:methods; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007211:Indoles; D007381:Intermediate Filament Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000502:physiology*; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D013234:Stem Cells / Q000502:physiology*; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection / Q000379:methods; D014404:Tubulin / Q000378:metabolism
D004268:DNA-Binding Proteins; D007211:Indoles; D007381:Intermediate Filament Proteins; C577484:NES protein, human; C539882:NR4A2 protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D014157:Transcription Factors; D014404:Tubulin; D049452:Green Fluorescent Proteins; C007293:DAPI; D004247:DNA; D001973:Bromodeoxyuridine