大鼠脑脊神经元Cell Specification
The tissue of the central nervous system is made up of two classes of cells that may be broadly
categorized as neurons and glia. Neurons are anatomic, functional, and trophic units of the brain
[1]. Despite great variability in size and shape, all neurons share common morphological
features, which are those of the key elements of a highly complex communication network. The
neurons are the dynamically polarized cells that serve as the major signaling unit of the nervous
system.
The RN-r from ScienCell Research Laboratories are isolated from E14 rat brain striatum. RN-s
are cryopreserved at primary cultures and delivered frozen. Each vial contains >1 x 106
cells in 1
ml volume. RN-r are characterized by immunofluorescent method with antibodies to
neurafilament, MAP2, and beta-tubulin 3. RN-r are negative for mycoplasma, bacteria, yeast and
大鼠脑脊神经元fungi. RN-r are guaranteed to further culture in the conditions provided by ScienCell Research
Laboratories.
Recommended Medium
It is recommended to use Neuronal Medium (NM, Cat. No. 1521) for the culture of rat neurons
in vitro.
Product Use
RN-r are for research use only. They are not approved for human or animal use, or for
application in in vitro diagnostic procedures.
Storage
Transfer cells directly and immediately from dry ice to liquid nitrogen upon receiving and keep
the cells in liquid nitrogen until cell culture is needed for experiments.
Shipping
大鼠脑脊神经元Dry ice.
Reference
[1] Parent, A. (1996) Neurons in Carpenters Human Neuroanatomy. 9th ed., pp131-198,
Williams & Wilkins, Quebec, Canada.
[2] Alberts, B., Bray, D., Lewis, J., Raff, M., Roberts, M., Watson, J. D. (1989) Molecular
biology of the cell. 2nd. ed., New York: Garland.
ScienCell
Research Laboratories
TM
Instruction for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37oC water bath
and return them to culture as quickly as possible with minimal handling!
Set up culture after receiving the ordering:
1. Prepare a poly-L-lysine coated flask (2 μg/cm2
, T-25 flask is recommended). Add
5 ml of sterile water to a T-25 flask and then add 5 μl of poly-L-lysine stock solution (10
mg/ml, ScienCell cat. no. 0413). Leave the flask in incubator overnight (minimum one
hour at 37oC incubator).
2. Prepare complete medium: decontaminate the external surfaces of medium and
medium supplements with 70% ethanol and transfer them to sterile field. Aseptically
open each supplement tube and add them to the basal medium with a pipette. Rinse each
tube with medium to recover the entire volume.
3. Rinse the poly-L-lysine coated flask with sterile water twice and add 7 ml of
complete medium to the flask. Leave the flask in the hood and go to thaw the cells.
4. Place the vial in a 37oC water bath, hold and rotate the vial gently until the
contents are completely thawed. Remove the vial from the water bath immediately, wipe
it dry, rinse the vial with 70% ethanol and transfer it to a sterile field. Remove the cap,
being careful not to touch the interior threads with fingers. Using 1 ml Eppendorf pipette
gently resuspend the contents of the vial.
5. Dispense the contents of the vial into the equilibrated, poly-L-lysine coated
culture vessels. A seeding density of 20,000 cells/cm2
is recommended.
Note: Dilution and centrifugation of cells after thawing are not recommended since these
actions are more harmful to the cells than the effect of DMSO residue in the culture. It is
also important that neurons are plated in poly-L-lysine coated culture vessels that
promote cell attachment.
6. Replace the cap or cover, and gently rock the vessel to distribute the cells evenly.
Loosen cap if necessary to permit gas exchange.
7. Return the culture vessels to the incubator.
8. For best result, do not disturb the culture for at least 16 hours after the culture has
been initiated. Change the growth medium the next day to remove the residual DMSO
and unattached cells, then every other day thereafter.
Maintenance of Culture:
大鼠脑脊神经元1. Change the medium to fresh supplemented medium the next morning after establishing a
culture from cryopreserved cells.
2. Change the medium every two to three days thereafter.
It is not recommended that neurons be subcultured beyond their initial plating.
Caution: Handling animal derived products is potentially bioharzadous. Always wear gloves and safety glasses
when working these materials. Never mouth pipette. We recommend following the universal procedures for
handling products of animal origin as the minimum precaution against contamination [1].
[1]. Grizzle, W. E., and Polt, S. S. (1988) Guidelines to avoid personal contamination by infective agents in research
laboratories that use human tissues. J Tissue Culture Methods. 11(4).