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C8-D1A [Astrocyte type I clone]

規(guī)格:

價(jià)格:

編號(hào):B164102

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

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產(chǎn)品名稱(chēng)

C8-D1A [Astrocyte type I clone]

商品貨號(hào)

B164102

Organism

Mus musculus, mouse

Tissue

brain, cerebellum

培養(yǎng)基

Cell Type

Astrocyte

Product Format

frozen

Morphology

neuronal

Culture Properties

adherent

Biosafety Level

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

8 days

Strain

C57BL/6

Storage Conditions

liquid nitrogen vapor phase

Karyotype

pseudodiploid, pseudodiploid Ref

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

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.

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 ATCC CRL-2535 and the astrocyte type III cloned cell line named C8-D30 is available as ATCC CRL-2534.

One clone with microglial properties named C8-B4 is available as ATCC 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).

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%.

Subculturing

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

Remove and discard culture medium.
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.
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.
Add appropriate aliquots of the cell suspension to new culture vessels.
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.

Cryopreservation

Freeze medium: Complete growth medium 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Culture Conditions

Temperature: 37°C

Atmosphere: air, 95%; carbon dioxide (CO₂), 5%

Name of Depositor

B Pessac, D Trisler

Deposited As

mouse

References

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

Alliot F, et al. A spontaneously immortalized mouse microglial cell line expressing CD4. Brain Res. Dev. Brain Res. 95: 140-143, 1996. PubMed: 8873987

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

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