PMID- 19719965
OWN - NLM
STAT- MEDLINE
DCOM- 20091211
LR  - 20090901
IS  - 2542-5641 (Electronic)
IS  - 0366-6999 (Linking)
VI  - 122
IP  - 14
DP  - 2009 Jul 20
TI  - Study of brain-derived neurotrophic factor gene transgenic neural stem cells in 
      the rat retina.
PG  - 1642-9
AB  - BACKGROUND: Neural stem cells (NSCs) transplantation and gene therapy have been 
      widely investigated for treating the cerebullar and myelonic injuries, however, 
      studies on the ophthalmology are rare. The aim of this study was to investigate 
      the migration and differentiation of brain-derived neurotrophic factor (BDNF) 
      gene transgenic NSCs transplanted into the normal rat retinas. METHODS: NSCs were 
      cultured and purified in vitro and infected with recombinant retrovirus 
      pLXSN-BDNF and pLXSN respectively, to obtain the BDNF overexpressed NSCs 
      (BDNF-NSCs) and control cells (p-NSCs). The expression of BDNF genes in two 
      transgenic NSCs and untreated NSCs were measured by fluorescent quantitative 
      polymerase chain reaction (FQ-PCR) and enzyme-linked immunosorbent assay (ELISA). 
      BDNF-NSCs and NSCs were infected with adeno-associated viruses-enhanced green 
      fluorescent protein (AAV-EGFP) to track them in vivo and served as donor cells 
      for transplantation into the subretinal space of normal rat retinas, phosphated 
      buffer solution (PBS) served as pseudo transplantation for a negative control. 
      Survival, migration, and differentiation of donor cells in host retinas were 
      observed and analyzed with Heidelberg retina angiograph (HRA) and 
      immunohistochemistry, respectively. RESULTS: NSCs were purified successfully by 
      limiting dilution assay. The expression of BDNF gene in BDNF-NSCs was the highest 
      among three groups both at mRNA level tested by FQ-PCR (P < 0.05) and at protein 
      level measured by ELISA (P < 0.05), which showed that BDNF was overexpressed in 
      BDNF-NSCs. The results of HRA demonstrated that graft cells could survive well 
      and migrate into the host retinas, while the immunohistochemical analysis 
      revealed that transplanted BDNF-NSCs differentiated into neuron more efficiently 
      compared with the control NSCs 2 months after transplantation. CONCLUSIONS: The 
      seed cells of NSCs highly secreting BDNF were established. BDNF can promote NSCs 
      to migrate and differentiate into neural cells in the normal host retinas.
FAU - Zhou, Xue-mei
AU  - Zhou XM
AD  - Department of Ophthalmology, Second Affiliated Hospital of Harbin Medical 
      University, Heilongjiang, China.
FAU - Yuan, Hui-ping
AU  - Yuan HP
FAU - Wu, Dong-lai
AU  - Wu DL
FAU - Zhou, Xin-rong
AU  - Zhou XR
FAU - Sun, Da-wei
AU  - Sun DW
FAU - Li, Hong-yi
AU  - Li HY
FAU - Shao, Zheng-bo
AU  - Shao ZB
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Chin Med J (Engl)
JT  - Chinese medical journal
JID - 7513795
RN  - 0 (Brain-Derived Neurotrophic Factor)
SB  - IM
MH  - Animals
MH  - Brain-Derived Neurotrophic Factor/genetics/*metabolism
MH  - Cell Differentiation/physiology
MH  - Cell Movement/physiology
MH  - Cells, Cultured
MH  - Embryo, Mammalian/cytology
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Immunohistochemistry
MH  - Neurons/*cytology
MH  - Rats
MH  - Retina/*cytology/*metabolism
MH  - Stem Cell Transplantation
EDAT- 2009/09/02 06:00
MHDA- 2009/12/16 06:00
CRDT- 2009/09/02 09:00
PHST- 2009/09/02 09:00 [entrez]
PHST- 2009/09/02 06:00 [pubmed]
PHST- 2009/12/16 06:00 [medline]
PST - ppublish
SO  - Chin Med J (Engl). 2009 Jul 20;122(14):1642-9.