Journal
JOURNAL OF THE CHINESE MEDICAL ASSOCIATION
Volume 79, Issue 5, Pages 256-263Publisher
LIPPINCOTT WILLIAMS & WILKINS
DOI: 10.1016/j.jcma.2015.10.011
Keywords
basic fibroblast growth factor; brain-derived neurotrophic factor; cell culture; embryonic rat; neural stem cells; Panax notoginseng saponins
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Funding
- National Natural Science Foundation of China [81373830]
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Background: We aimed to study the effect of Panay notoginseng saponins (PNS) on the proliferation, differentiation, self-renewal, and expressions of basic fibroblast growth factor (bFGF) and brain-derived neurotrophic factor (BDNF) in rat embryonic neural stem cells (NSCs). Methods: Cortical stem cells were isolated from rat embryos on Embryonic Day 17 (E17) and identified by nestin expression. Subsequently, primary culture, subculturing, and single cell cloning were performed on the cells. After the first cell passage (P1), the cells were resuspended and divided into a control group and a treatment group. Control cells were cultured in serum-free basal culture medium with B27 and dulbecco's modified eagle medium (DMEM)/F12. The same medium supplemented with PNS (100 mu g/mL) was used to culture cells in the treatment group. Both groups were incubated at 37 degrees C in a 5% CO2 incubator. Immunocytochemistry was performed 4 days after incubation. Results: Primary, P1, and P2 cells in the treatment group formed neurospheres, as did single cell clones of the P1 cells in this group. After being cultured for 4 days, the number of nestin-, proliferating cell nuclear antigen (PCNA)-, Tuj-1-, neurofilament (NF)-, vimentin-, glial fibrillary acidic protein (GFAP)-, bFGF-, and BDNF-positive cells significantly increased in the treatment group in comparison to the control group. All positively stained cells could form clear clusters. Conclusion: PNS can promote rat embryonic cortical NSC survival, self-renewal, proliferation, and differentiation through neurotrophic factors by autocrine or paracrine signaling. Copyright (C) 2016, the Chinese Medical Association. Published by Elsevier Taiwan LLC.
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