Application of daucosterol in preparing medicines for promoting proliferation of neural stem cells
A technology of neural stem cells and carotene, applied in the field of medicine
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Embodiment 1
[0102] Embodiment 1. In Vitro Experiment
[0103] 1.1 Experimental animals
[0104] Ten pregnant SD rats at 14-16 days of pregnancy were provided by Zhejiang Experimental Animal Center, certificate number: SCXK (Zhejiang) 2008-0033.
[0105] 1.2 Main reagents and reagent preparation:
[0106] Neuralbasal medium (product of GIBCO company, Lot 1094141), B27 medium supplement (product of GIBCO company, Lot 1159693); rat b-FGF and EGF (product of Pepro TECH company, EGF: Lot 0906390-1 H2911); b-FGF: Lot 1305CY12) mouse anti-rat nestin monoclonal antibody, TRITC (rhodamine)-labeled goat anti-mouse IgG antibody, Brdu and its antibody (product of Wuhan Boster Biotechnology Co., Ltd.); CCK-8 kit (Beyond Institute of Biotechnology); Carotin, provided by Jiangsu Provincial Key Laboratory of Prescription Research, Nanjing University of Traditional Chinese Medicine, purity ≥ 98%;
[0107] Dissecting fluid:D-hank , s, (pH7.2-7.4): NaCl8.0g, Na 2 HP0 4 12H 2 O0.132g, KH 2 PO 4...
Embodiment 2
[0143] Embodiment 2. overall experiment
[0144] 2.1 Animals, grouping and handling
[0145] Twenty aged mice (11 months old), half male and half male, were divided into blank control group and carrotin group, 10 mice in each group. The blank control group was administered with normal saline, and the carotin group was administered with 10 mg / kg carotin, once a day for 30 days.
[0146] 2.2 Brdu-nestin double labeling and laser confocal microscope photography
[0147] method: Three days before the animals were sacrificed, the mice were given intraperitoneal injection of Brdu, 50 mg / kg, once every 12 hours, for three consecutive days. 12 hours after the end, the mice were anesthetized by intraperitoneal injection of chloral hydrate, the blood vessels were flushed with normal saline, and the heart was perfused with 4% paraformaldehyde. The brain was quickly fixed and removed, and fixed in 4% frozen paraformaldehyde for 48 hours. Then dehydrate in 20%, 30% sucrose hype...
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