宁波泰斯拓生物

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C8-D1A [Astrocyte type I clone]

货号 TS212512
中文名称 null
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产品名称: C8-D1A Astrocyte type I clone
商品货号: TS212512
Organism: Mus musculus, mouse
Tissue: brain, cerebellum
Cell Type: Astrocyte
Product Format: frozen
Morphology: neuronal
Culture Properties: adherent
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: 8 days
Strain: C57BL/6
Storage Conditions: liquid nitrogen vapor phase
Karyotype: pseudodiploid, pseudodiploid RefAlliot 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 Dulbeccos Modified Eagles 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.

  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. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. 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