Method for inducing SH-SY5Y cells to differentiate into dopaminergic neurons
A SH-SY5Y, dopaminergic technology, applied in the field of neuroscience research, can solve the problem of lack of cell models, and achieve the effect of high differentiation level and high differentiation rate
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0026] Example 1 Culture medium preparation
[0027] Basal medium: DMEM / F12 medium (Gibco), FBS (15%, Gibco), Antibiotic-Antimycotic mixture (1%, Gibco).
[0028] Induction medium S1: DMEM / F12 medium (Gibco), B27 (1:50, Invitrogen), FBS (1%, Gibco), Antibiotic-Antimycotic mixture 1%, heparin sodium (5 μg / ml, Changzhou Qianhong Biochemical Co., Ltd. ), L-glutamine (2 mM, Invitrogen), aFGF (100 ng / mL, R&D), TPA (phorbol 12-myristate 13-acetate, 100 nM, sigma), forskolin (25 μM, sigma), dbcAMP ( 100 μM, sigma), pramipexole (10 μM, sigma).
[0029]Induction medium S2: DMEM / F12 medium (Gibco), B27 (1:50, Invitrogen), FBS (1%, Gibco), Antibiotic-Antimycotic mixture 1%, heparin sodium (5 μg / ml, Changzhou Qianhong Biochemical Co., Ltd. ), L-glutamine (2 mM, Invitrogen), aFGF (50 ng / mL, R&D), TPA (phorbol 12-myristate 13-acetate, 50 nM, sigma), forskolin (25 μM, sigma), dbcAMP ( 50 μM, sigma), pramipexole (10 μM, sigma).
[0030] Induction medium S3: DMEM / F12 medium (Gibco), B27 (1...
Embodiment 2
[0032] Example 2 Culture of SH-SY5Y cells
[0033] Recovery: Take the purchased SH-SY5Y cells (purchased from Nanjing Kaiji Biotech) 1×10 6 In 10mL basal medium, centrifuge at 1000rpm for 5min, discard the supernatant. Take 8mL basal medium to resuspend the cells, transfer to a T25 cell culture flask, and place at 37°C, 5% CO 2 cultured in an incubator.
[0034] Cultivation: The cell culture medium was changed every 2 days.
[0035] Subculture: When the cell adhesion reaches more than 80%, proceed to subculture. Carefully aspirate the medium, add 10 mL of PBS (Hyclone) buffer to wash the cells once, and discard the buffer. Add 1 mL of Accutase (StemCell TECHNOLOGIES) to each T25 flask of cells, and digest at 37°C for 1-2 min until the cells are completely detached from the culture flask. 4 mL of basal medium terminated the digestion. Transfer the digested cell suspension into a 15mL centrifuge tube, centrifuge at 1000rpm for 5min, discard the supernatant, resuspend the c...
Embodiment 3
[0036] Example 3 Induced differentiation of SH-SY5Y cells
[0037] The SH-SY5Y cells subcultured in Example 2 were cultured in basal medium, and when the cell confluence reached 80%, the cells were digested with Accutase. Place 4 polylysine-coated glass slides (cell slides) in a Φ60mm petri dish, and add 1×10 5 For each SH-SY5Y cell, the basal medium supplemented the system to 4mL. After the SH-SY5Y cells adhered to the wall after culturing for 24 hours, all the basal medium was discarded and replaced with 4 mL of induction medium S1. The induction medium S1 was completely changed every 2 days, and the dopaminergic neurons were formed after 7 days of culture.
PUM
Property | Measurement | Unit |
---|---|---|
diameter | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com