小鼠小胶质细胞Cell Specification
Microglia, one of the glial cell types in the CNS, is an important integral component of the
neuro-glial cell network [1]. They have been observed in the brain parenchyma from the early
stage of development to the mature state. Microglia act as brain macrophages when programmed
cell death occurs during brain development or when the CNS is injured or pathologically
damaged. Microglia can be considered the main cell in brain immune surveillance, can present
antigens in the molecular context of MHC class II expression to CD-4 positive T cells, are
capable of Fc-mediated phagocytosis, and share many common antigens with hemopoietic and
tissue macrophages [2]. Furthermore, there is accumulating evidence that microglia are involved
in a variety of physiological and pathological processes in the brain by interacting with neurons
and other glial cells and through production of biologically active substances such as growth
factors, cytokines, and other factors [3].
MM from ScienCell Research Laboratories are isolated from neonate day two C57BL/6 mouse
brain tissue. Cells are harvested after purification and delivered frozen. Each vial contains >1 x
10^6 cells in 1 ml volume. MM is characterized by immunofluorescent method with antibody to
F4/80. MM is negative mycoplasma, bacteria, yeast and fungi. M is not recommended for
expanding or long term cultures since the cells do not proliferate in culture.
小鼠小胶质细胞Recommended Medium
It is recommended to use Microglia Medium (MM, Cat. No. 1901) for the culturing of MM in
vitro.
Product Use
MM is 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 or gel ice.
Reference
[1] Lee, S. C., Liu, W., Brosnan, C. F. and Dickson, D. W. (1992) Characterization of primary human fetal
dissociated central nervous system cultures with an emphasis on microgia. Laboratory Investigation. 67:465-476.
[2] Fedoroff, S., Zhai, R. and Novak, J. P. (1997) Microglia and astroglia have a common progenitor cell. J.
Neurosci. Res. 50: 477-486.
[3] Stoll, G. and Jander, S. (1999) The role of microglia and macrophages in the pathophysiology of the CNS. Prog.
小鼠小胶质细胞Neurobiol. 58:233-247.
Instruction for culturing cells
Caution: Cryopreserved cells are very delicate. Thaw the vial in a 37oC waterbath
and return them to culture as quickly as possible with minimal handling!
Set up culture after receiving the order:
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 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.
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 ≥10,000 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 the cells are plated in poly lysine coated culture vessels that
promote the cell attachment.
6. Replace the cap or cover, and gently rock the vessel to distribute the cells evenly. Loosen
小鼠小胶质细胞caps 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. A health culture of microglia show
characteristic elongated, almost bipolar cell bodies with spine-like processes that often
branch perpendicularly.
Maintenance of Culture:
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 microglia be subcultured beyond their initial plating.
小鼠小胶质细胞Caution: Handling animal derived products is potentially biohaza dous. 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).