C8-B4細胞
細胞形態： 神經元
生長狀態： 貼壁生長
ATCC Number： CRL-2540?
運輸方式： 凍存運輸
年限： 8 days juvenile
細胞類型： 其他細胞類型
是否是腫瘤細胞： 0
物種來源： 小鼠
品系： C57BL/6
組織來源： cerebellum
器官來源： 大腦
數量： 大量
C8-B4細胞Designations: C8-B4
Depositors: B Pessac, D Trisler
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Mus musculus
Morphology: neuronal
Source: Organ: brain
Strain: C57BL/6
Tissue: cerebellum
Cell Type: monocyte/macrophage microglia; spontaneously transformed
Cellular Products: glutamate [49701]
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Antigen Expression: CD4 [49701]
Age: 8 days juvenile
Comments: Clonal permanent cell lines with astrocytic or microglial properties have been established from explant cultures of 8-day postnatal mouse cerebella after in vitro spontaneous transformation. [48963] [49701]
Sister flasks of lethally irradiated astrocytes, which are known to synthesize the macrophage-microglia growth factor, M-CSF, were used as a substrate for cloning the cells. [48963] [49701]
The cell lines were derived in a multistage process. Slowly proliferating foci with several morphologies appeared 4 months after initiation of the cultures and became progressively enriched by cells with a homogeneous appearance. [48963]
The astrocyte type I cloned cell line named C8-D1A is available as ATCC CRL-2541, the astrocyte type II cloned cell line named C8-S is available as ATCC CRL-2535 and the astrocyte type III cloned cell line named C8-D30 is available as ATCC CRL-2534.
The C8-B4 clone expresses classical microglial markers (MAC1, F4/80, 2-4G2) and appears to be derived from a committed microglial precursor since it does not express differentiation antigens present during the early stage of the monocytic lineage. [49701]
C8-B4 cells synthesize the CD4 molecule and produce and release large amounts of glutamate. [49701]
Propagation: C8-B4細胞ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Temperature: 37.0°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Subculturing: Protocol: NOTE: The cells do not become confluent.
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and 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.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to10 minutes.Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
Place culture vessels in incubators at 37?C. Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37?C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 48 to 72 hrs
Related Products: C8-B4細胞Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
purified RNA:ATCC CRL-2540R
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
References: 48963: Alliot F, Pessac B. Astrocytic cell clones derived from established cultures of 8-day postnatal mouse cerebella. Brain Res. 306: 283-291, 1984. PubMed: 6466977
49701: Alliot F, et al. A spontaneously immortalized mouse microglial cell line expressing CD4. Brain Res. Dev. Brain Res. 95: 140-143, 1996. PubMed: 8873987
