Neural restoration promoting material as well as preparation method and application thereof
A nerve repair and nerve growth factor technology, applied in the field of nerve repair materials, can solve the problems of unsatisfactory drug release stability, difficult control of release speed and activity, and potential safety hazards, so as to achieve safe use and stable effect , low-cost effect
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Embodiment 1
[0043] Embodiment 1 Preparation and detection of nerve-promoting repair materials of the present invention
[0044] 1. Isolation, purification, culture and detection of Schwann cell SCs
[0045] 1. Isolation, purification and culture of Schwann cell SCs
[0046] (1) Material collection: 20 SD suckling mice aged 5-7 days were taken and killed by necking. Under sterile conditions, the bilateral sciatic nerves were taken, and the epineurium tissue was peeled off as soon as possible under a dissecting microscope with microscopic instruments.
[0047] (2) Digestion: Digest with 0.25% trypsin and 0.2% type II collagenase for 10 minutes respectively, centrifuge at 1200rpm for 5 minutes, and inoculate once for 20 minutes after differential attachment.
[0048] (3) Purification: After 24 hours, replace with 20μl / ml G418 (200μg / mL) or 2×10 -6 M~1×10 -5 M cytarabine was cultured in 10% FBS H-DMEM medium. After two days of G418 or cytarabine treatment, the H-DMEM medium containing 10%...
Embodiment 2
[0090] Example 2 Preparation and detection of nerve-promoting repair materials of the present invention
[0091] 1. Subculture, cryopreservation and detection of RSC96 cells
[0092] 1. Subculture of RSC96 cells
[0093] (1) Subculture: RSC96 cells were purchased from the Cell Bank of the Chinese Academy of Sciences. Such as Figure 7 When the RSC96 cells grow to about 80% confluent, aspirate the medium, add 0.5ml of PBS solution containing 0.25% trypsin and 0.1% EDTA to digest, shake the digestion solution for about 2-5 minutes, the cells shrink and change under the microscope After the circle or a few fall off, gently shake the bottom of the bottle to make all the cells fall off, add 5ml of H-DMEM medium containing 10% FBS, blow gently to make the cells basically form a single suspension, and divide them into 4 cell culture flasks. Supplement H-DMEM medium containing 10% FBS to about 5ml, place in 5% CO 2 cultured in a constant temperature incubator at 37°C.
[0094] (2...
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