C8-D1A astrocyte cell line小鼠脑星形胶质神经元细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

C8-D1A astrocyte cell line小鼠脑星形胶质神经元细胞株

C8-D1A [Astrocyte type I clone] Cell Line
Cat No.: BioVector-A2541

C8-D1A [Astrocyte type I clone] cells are astrocytes isolated from the cerebellum of mice. Use them in neuroscience research applications.
Product category
Animal cells
Organism
Mus musculus, mouse
Classification
Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Cell type
astrocyte
Morphology
neuronal
Tissue
Brain; Cerebellum
Applications
3D cell culture
Neuroscience

Characteristics
Growth properties
Adherent
Derivation
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. 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.
Age
8 days
Strain
C57BL/6
Karyotype
pseudodiploid, pseudodiploid
Comments
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.

Some of these cloned cell lines bound anti-glial fibrillary acidic protein (GFAP) antibodies and therefore appeared to be astrocytic. No other glial neuronal or microglial markers have been detected in these clones.
According to their morphology, 3 separate types of these GFAP-positive clones could be distinguished. Type I and II cells had small somata.
Type I had several short processes, while type II had two processes, one of which was very thin and long (greater than 200 microns). Type III cells had large flat somata and no processes.
The astrocyte type II cloned cell line named C8-S is available as BioVector-A2535 and the astrocyte type III cloned cell line named C8-D30 is available as BioVector-A2534.
One clone with microglial properties named C8-B4 is available as BioVector CRL-2540.
The C8-D1A cell line has the morphology of fibrous astrocytes.
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.
These astrocytic clones might be the in vitro counterparts of fibrous (type I), or velamentous (type III) astrocytes and of Golgi epithelial cells (type II).

Handling information
Unpacking and storage instructions
1.Check all containers for leakage or breakage.
2.Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below _x001f_-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
The base medium for this cell line is BioVector-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°C
Atmosphere
95% Air, 5% CO2
Handling procedure
To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
1.Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
2.Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3.Transfer the vial contents to a 75 cm2 tissue culture flask and dilute with the recommended complete culture medium (see the specific batch information for the recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
4.Incubate the culture at 37°C in a suitable incubator. A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

Note: If it is desired that the cryoprotective agent be removed immediately, or that a more concentrated cell suspension be obtained, centrifuge the cell suspension at approximately 125 x g for 5 to 10 minutes. Discard the supernatant and resuspend the cells with fresh growth medium at the dilution ratio recommended in the specific batch information.
Subculturing procedure
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
1.Remove and discard culture medium.
2.Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53% (w/v) EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3.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.
4.
5.Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
6.Add appropriate aliquots of the cell suspension to new culture vessels.
7.Incubate cultures at 37°C

Subculture Ratio: 1:4 to 1:6
Medium Renewal: Every 2 to 3 days.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO

Quality control specifications
Mycoplasma contamination
Not detected

BSL：Level 1

Supplier来源:BioVector NTCC Inc.
TEL电话:400-800-2947
Website网址: http://www.biovector.net