Revisiting nestin expression in retinal progenitor cells in vitro and after transplantation in vivo.
The purpose of this study is to characterize the co-expression of nestin--a neuroectodermal stem cell and a reactive glial marker-with various mature retinal cell markers in retinal progenitor cells (RPCs) expanded in vitro, followed either by in vitro induction or subretinal transplantation. Rat RPCs derived from embryonic day (E) 17 rat retina were expanded in serum free defined culture, and induced to differentiate by all-trans retinoic acid (RA). Following induction, cells were stained for nestin in combination with retinal neuronal and glial markers. Cultured cells were collected for quantitative RT-PCR gene expression analysis prior to and after induction. In a second series, passage 2 RPCs were transplanted into the subretinal space of S334ter-3 retinal degeneration rats at postnatal day 28. After 1-4 weeks, sections through the transplant were double immunostained for nestin and various retinal specific neuronal markers. The cultured RPCs treated with RA exhibited nestin co-expression with various retinal specific markers, including protein kinase C alpha (PKC), neurofilament 200 (NF200), cellular retinaldehyde binding protein (CRALBP), and rhodopsin. Following RA induction, quantitative RT-PCR analysis demonstrated downregulation of nestin, PAX-6, thy1.1, and PKCalpha, and upregulation of rhodopsin, glial fibrillary acidic protein (GFAP), and CrX. No nestin coexpression was observed with any of the retinal specific neuronal markers in RPC transplants in vivo except for some nestin-immunoreactivity overlapping with GFAP positive cells in the host retina. The role of nestin as a unique neural stem/progenitor cell marker should be reconsidered. Nestin expression during RPC maturation appears to be different in vitro versus in vivo.
Qiu
Guanting
G
Department of Ophthalmology, Doheny Retina Institute, Keck School of Medicine, at the University of Southern California, 1450 Pablo St - DEI 3610, Los Angeles, CA, 90033-3699, USA.
Seiler
Magdalene J
MJ
Thomas
Biju B
BB
Wu
Kebin
K
Radosevich
Michael
M
Sadda
SriniVas R
SR
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D015536:Down-Regulation; D005260:Female; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D008297:Male; D008858:Microscopy, Phase-Contrast; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D064231:Nestin; D051381:Rats; D011922:Rats, Mutant Strains; D012160:Retina / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D012162:Retinal Degeneration / Q000378:metabolism / Q000628:therapy*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D014212:Tretinoin