CD1小鼠海马趾神经元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, the key elements of a highly complex communication network. Neurons are
dynamically polarized cells that serve as the major signaling unit of the nervous system. The
brain is known to contain billions of neurons, each being able to contact thousands of other
neurons [2].
MN-h from ScienCell Research Laboratories are isolated from embryonic day 18 mouse
hippocampus. MN-h are cryopreserved as primary cultures and delivered frozen. Each vial
contains >1 x 106
cells in 1 ml volume. MN-h are characterized by immunofluorescence with
antibodies specific to neurofilament, MAP2, and β-tubulin III. MN-h are negative for
CD1小鼠海马趾神经元mycoplasma, bacteria, yeast, and fungi. MN-h are guaranteed to further culture under the
conditions provided by ScienCell Research Laboratories; however, MN are not recommended
for expanding or long term cultures since the cells do not proliferate in culture.
Recommended Medium
It is recommended to use neuronal medium (NM, Cat. #1521) for culturing MN-h in vitro.
Product Use
MN-h are for research use only. They are not approved for human or animal use, or for
application in in vitro diagnostic procedures.
Storage
Upon receiving, directly and immediately transfer the cells from dry ice to liquid nitrogen and
keep the cells in liquid nitrogen until they are needed for experiments.
Shipping
Dry ice.
References
[1] Parent A. (1996) “Neurons.” In Carpenters Human Neuroanatomy (9th ed., pp131-198). Quebec: Williams &
Wilkins.
[2] Alberts B, Bray D, Lewis J, Raff M, Roberts M, Watson JD. (1989) Molecular Biology of the Cell (2nd ed.).
New York: Garland.
Instructions for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37oC water bath
and return the cells to culture as quickly as possible with minimal handling!
Initiating the culture:
1. Prepare a poly-L-lysine-coated culture vessel (2 μg/cm2
). It is recommended to use a T-25
CD1小鼠海马趾神经元flask (alternatively, 3 wells of a 6-well plate or 12 wells of a 24-well plate can be used).
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, Cat. #0413). Leave the vessel in a 37oC incubator overnight (or for a
minimum of one hour). Rinse the poly-L-lysine-coated vessel twice with sterile water
prior to use.
Note: It is important that these cells are plated in poly lysine coated culture vessels to
promote cell attachment.
2. Prepare complete medium. Decontaminate the external surfaces of medium bottle and
medium supplement tubes with 70% ethanol and transfer them to a sterile field.
Aseptically transfer supplement to the basal medium with a pipette. Rinse the supplement
tube with medium to recover the entire volume.
3. Add complete medium to the culture vessel. Leave the vessel in the sterile field and
proceed to thaw the cryopreserved cells.
4. Place the frozen vial in a 37oC water bath. Hold and rotate the vial gently until the
contents completely thaw. Promptly remove the vial from the water bath, wipe it down
with 70% ethanol, and transfer it to the sterile field. Carefully remove the cap without
touching the interior threads.
5. Gently resuspend and dispense the contents of the vial into the poly-L-lysine-coated
culture vessel. A seeding density of 10,000-50,000 cells/cm2
is recommended, with an
optimal range of 20,000-25,000 cells/cm2
.
Note: Dilution and centrifugation of cells after thawing are not recommended since these
actions are more harmful to the cells than the effect of residual DMSO in the culture.
6. Replace the cap or lid of the culture vessel and gently rock the vessel to distribute the
cells evenly. Loosen cap, if necessary, to allow gas exchange.
CD1小鼠海马趾神经元7. Return the culture vessel to the incubator.
8. For best results, do not disturb the culture for at least 16 hours after the culture has been
initiated. Refresh culture medium the next day to remove residual DMSO and unattached
cells, then every other day thereafter.
Maintaining the culture:
1. Refresh supplemented culture 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 biohazardous. Always wear gloves
and safety glasses when working with 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 WE, Polt S. (1988) “Guidelines to avoid personal contamination by infective agents in research
laboratories that use human tissues.” J Tissue Cult Methods. 11: 191-9.