Rat cerebral cortical neuron cell culture can: (1) obtain rat cerebral cortical neurons; (2) for neuronal cell differentiation; (3) for neuronal apoptosis. We are the one of three leading factory in China, specialized in manufacturing laser distance module since 2004. Bilateral laser distance measurer is one of mutifunctional laser distance device. JRT multifunctional laser measuring device functions include area,distance,length,volume,continuous measurement,addition and subtraction, measuring unit: ft/m/inch. More functions can be customerized. And the laser measuring meter have distance 30/40/60/80/100/150m for chosing. This bilateral degin is our 40m middle laser distance transducer 703A laser measuring meter. Laser Range Finder Measurer,80M Laser Distance Meter,Distance Meter 80M,Laser Distance Meters Chengdu JRT Meter Technology Co., Ltd , https://www.infrareddistancesensor.com experimental method
Principle of experimental method The SD rat fetal rat cortical neurons were cultured in vitro for 7 days. The micropipette plastic dripper was mechanically cut into the cultured neurons. The neurons were divided into light, medium and heavy groups according to the degree of cutting. No mechanical scribing was performed, and the rest of the treatment was the same as the injury group. Cell viability and culture supernatant supernatant lactate dehydrogenase (LDH) were measured at different time points (10, 30 min, 1, 3, 6, 12, 24 h) after injury. content. Experimental Materials Reagents, kits Instruments, consumables Experimental procedure Precautions
OEM service:
* usb chargable
* voice
* IP54 rating
* Colorful case
Cell Technology Topic: Rat Cerebral Cortical Neuron Cell Culture Experiment
SD rat
Borate Borax Buffer Trypsin-EDTA Digestion D-Hanks'
Ophthalmic scissors dropper centrifuge Petri dish CO2 incubator
First, the experimental steps
1. 16 d pregnant SD rats were anesthetized with pentobarbital sodium, iodine, 75% ethanol disinfected abdominal skin, cut skin, muscle, subcutaneous fascia in turn, aseptically removed embryos into pre-cooled D -Hanks' balanced salt solution.
2. Separate and remove the embryonic cerebral cortex under a dissecting microscope, remove the meninges, cut the tissue into a size of about 1 mm 3 , add the trypsin-EDTA digest and place in a 37 ° C incubator for 20 min, shaking once in the middle.
3. The tissue was then aspirated with a dropper and transferred to a centrifuge tube containing pre-chilled culture medium (DMEM + 10% FBS) to terminate the digestive juice for 5 min.
4. Aspirate the tissue with a dropper and transfer it to a centrifuge tube containing pre-cooled DMEM + 10% FBS. Blow it several times with a flame-polished Pasteur pipette. After standing, take the supernatant and pipette it into another centrifuge tube. Repeat the above steps 2 to 3 times.
5. Filter the collected supernatant through a 200 mesh screen.
6. The filtered cell suspension was centrifuged at 800 rpm for 5 min, the supernatant was discarded, fresh medium (DMEM + 20% FBS) was added to the tube and pipetted into a single cell suspension.
7. Count the cells, adjust the cell density into 1.5-well plates at 1.5 × 10 5 /cm 2 , and incubate them in a CO 2 incubator. After 48 h of culture, the solution was changed and Ara-C was added to give a final concentration of 1 × 10 -5 mM / L. After Ara-C was applied for 24 h, the whole amount of liquid was changed. Change the liquid once every three and a half days later.
1. The above steps are traditional methods. There are many places that can be modified: for example, it can be directly digested with 0.125-0.25% trypsin for 15-20 min during digestion. It can also be digested with DNase. Some people can also directly blow it.
2. Although the rat cortical neurons are above, they are also suitable for mice, but mice of about 13 days of pregnancy should be selected.
3. Neurons are a relatively difficult cell to grow, so the density of cells must be sufficient at the time of inoculation.
4. When cultivating neurons on slides, it is important to pay attention to the source and treatment of the slides. This is very important and has a great impact on nerve cells. If the neurons you are growing on the slides are still pretty good, then you should use slides from the same source (manufacturer) in future cultures.
5. If B27 and neurobasal medium are available, it is convenient (without adding AraC):
(1) Suspension of the cell suspension (DMEM + 20% FBS), and after 6-12 h on the culture dish, the whole amount was changed to 2% B27 neurobasal medium, and the culture was continued, and the medium was changed for 3 days.
(2) The cell suspension was inoculated by suspension directly with 2% B27 of neurobasal medium.
(3) It can be directly cultured in the fetal rat cortex within 1 day of newborn.