大鼠黑质神经元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 morphologic 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 human brain is known to contain about 1 x 1011 neurons, each being able to contact at least
10,000 other neurons [2].
RN-c from ScienCell Research Laboratories were isolated from E18 rat brain cortical. RN-c
were cryopreserved at primary culture and delivered frozen. Each vial contains >1 x 10 cells in
1 ml volume. RN-c are characterized by immunofluorescent method with antibodies to
neurafilament, MAP2, and beta-tubulin III. RN-c are negative for mycoplasma, bacteria, yeast
and fungi. RN-c are guaranteed to further culture in the conditions provided by ScienCell
Research Laboratories.
Recommended Medium
大鼠黑质神经元It is recommended to use neuronal medium (NM, Cat. No. 1521) for the culturing of RN-c in
vitro.
Product Use
RN-c 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
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.
Instruction for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37 C waterbath
and return them to culture as quickly as possible with minimal handling!
Unpacking
1. For crypreserved cells: If there is dry ice in the package and you are not going to culture
cells right way, place cryovial(s) immediately into liquid nitrogen. If there is no dry ice
left in the package, thaw and culture the cells immediately.
2. For proliferating cells: Spray the culture vessel (flask, plate or slide) with 70% ethanol
for disinfection. Transfer the cells into 37 C, 5% CO incubator and allow equilibrating
for 2 hours. After cells have equilibrated, remove shipping medium from the culture
vessel and replace with fresh medium.
Set up culture after receiving the ordering
1. Coat culture vessel with laminin or poly-L-lysine.
Note: It is important that neurons are plated in laminin or poly-L-lysine coated culture
vessels that promote cell attachment and neurites outgrowth (poly-L-lysine coating: coat
flask or plate with poly-L-lysine at 2 μg/ml concentration for one hour and wash the flask
or plate with sterile water three times).
大鼠黑质神经元2. Medium preparation: 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
the medium to recover the entire volume.
3. Set up culture: Prepare one T-45 flask for each cryovial. Add the appropriate amount of
medium to the vessel (recommend for 10 ml/T-45 flask) and allow the flask to equilibrate
in 37 C, 5% CO incubator for at least 30 min.
4. Thawing of cells: Place the vial in a 37 C waterbath, hold and rotate the vial gently until
the contents are completely thawed. Remove the vial from the waterbath immediately,
wipe it dry, and transfer it to a sterile field. Rinse the vial with 70% ethanol, and then
wipe to remove excess. Remove the cap, being careful not to touch the interior threads
with fingers.
5. Using 1 ml eppendorf pipette gently resuspend the cells in the vial and transfer them to
equilibrated culture vessels (a T-45 flask). A high seeding density (>10,000/cm ) 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.
6. Replace the cap or cover, and gently rock the vessel to distribute the cells evenly. Loosen
caps if necessary to permit gas exchange. Return the culture vessels to the incubator.
7. Change the medium 12 hours after plating to remove the residual DMSO and unattached
cells, then every other day thereafter. A health culture will display normal neuron
morphology, and nonvacuole cytoplasm with multiple processes.
大鼠黑质神经元Caution: Handling human derived products is potentially bioharzadous. Although each cell strain testes negative
for HIV, HBV and HCV DNA, proper precautions mush be taken to avoid inadvertent exposure. Always wear gloves
and safety glasses when working these materials. Never mouth pipette. We recommend following the universal
procedures for handling products of human 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).