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SIM-A9
SIM-A9
規(guī)格:
價格:
編號:B238164
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱
SIM-A9
商品貨號
B238164
Organism
Mus musculus, mouse
Tissue
brain; cerebral cortex
培養(yǎng)基
Cell Type
microglia
Product Format
frozen 1.0 mL
Morphology
neuronal-like
Culture Properties
mixed, adherent with some single cells in suspension
Biosafety Level
1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Age
postnatal 1 day
Strain
C57BL/6
Applications
May serve as a model system for the investigation of microglial behavior in vitro in response to inflammatory stimuli, screening immunomodulating compounds, and brain malignancy research.
Storage Conditions
liquid nitrogen vapor phase
Images
Cell Micrograph of SIM-A9 Mouse Brain Microglia Cells
Derivation
This cell line was derived from cortical tissues collected and pooled from mouse pups at postnatal day 1. The tissues were dissociated by trypsinization and plated. On day 14 the microglial cells were harvested by vigorously shaking the flasks and the plating out detached microglial cells. The plates were maintained for 2 weeks at which time an unexpected, extensive proliferation of microglial cells were observed. The cells were passaged 7 times over a 4 week period, and were determined to have become spontaneously immortalized. The cells were then plated out in a limited dilution procedure and colonies derived from wells observed to contain a single cell were expanded. A clone designated as “A-9” gave rise to this cell line.
Antigen Expression
CD68+; ionizing calcium-binding adaptor molecule 1 (Iba1)+   RefNagamoto-Combs K, et al. A novel cell line from spontaneously immortalized murine microglia. J. Neurosci. Methods 233: 187-198, 2014. PubMed: 24975292
Comments
These spontaneously immortalized micgoglial cells exhibit phagocytic activity upon liposaccharide stimulation and TNFα cytokine secretion upon β-amyloid exposure. These cells may be used in place of primary microglial cultures, which require a lengthy and costly preparation procedure.
Complete Growth Medium
The base medium for this cell line is DMEM:F12 Medium Catalog No. 30-2006. To make the complete growth medium, add the following components to the base medium:
  • heat-inactivated horse serum to a final concentration of 5%
  • heat-inactivated fetal bovine serum to a final concentration of 10%

Subculturing

Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Transfer all medium and floating cells from flask to a centrifuge tube.
  2. Briefly rinse the cell layers with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS). Add rinse solution to the cells in centrifuge tube in step 1.
  3. Add 5.0 ml of phosphate buffered saline containing 1 mM EDTA, 1 mM EGTA and 1 mg/mL glucose solution to flask to detach cells. Observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. After the cells have detached, transfer them to the centrifuge tube in step 1.
  5. Centrifuge the cells at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium.
  7. Add appropriate aliquots of cell suspension to new culture vessels.
  8. Place culture vessels in incubators at 37°C.
Subcultivation Ratio: 1:3 to 1:6 

Medium renewal: every 2 to 3 days

Cryopreservation
Complete growth medium, 90%; DMSO, 10%
Culture Conditions
Temperature: 37°C
Volume
1.0 mL
Name of Depositor
K Nagamoto-Combs
References

Nagamoto-Combs K, et al. A novel cell line from spontaneously immortalized murine microglia. J. Neurosci. Methods 233: 187-198, 2014. PubMed: 24975292

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