AtT-20/D16v-F2細胞

是否是腫瘤細胞： 0

物種來源： 小鼠

數(shù)量： 大量

生長狀態(tài)： 貼壁生長

品系： LAF1

器官來源： 其他

運輸方式： 凍存運輸

ATCC Number： CRL-1795?

相關(guān)**： 腫瘤

細胞形態(tài)： 其他

AtT-20/D16v-F2細胞Designations: AtT-20/D16v-F2

Depositors: RB Kelly

Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: loosely adherent

Organism: Mus musculus

Morphology: small rounded cells

Source: Organ: pituitary, anterior

Strain: LAF1

Disease: tumor

Cellular Products: adrenocorticotropic hormone (ACTH)

Permits/Forms: AtT-20/D16v-F2細胞In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Comments: The F2 subclone of AtT-20 (see ATCC CCL-89) was developed by B. Gumbiner.

This clone has been been used successfully by Moore et al. for several DNA mediated transfection studies relating to endocrine and exocrine secretory pathways.

The cells are readily transfected using a standard calcium phosphate protocol.

Tested and found negative for ectromelia virus (mousepox).

Propagation: 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%.

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

Temperature: 37.0°C

Subculturing: Protocol:

Cells grow in patches and pile up. Cultures do not become confluent.

AtT-20/D16v-F2細胞Volumes used in this protocol are for 75 sq cm flasks; 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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 minutes).

Note: Extreme care must be taken to prevent the cells from clumping. Carefully follow the subcultivation protocol .do not over trypsinize. The cells will slough if the pH of the medium becomes too acidic (lower than 7.5).

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Transfer cell suspension to a centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes. Discard supernatant.

Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.

Incubate cultures at 37C.

Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended

Medium Renewal: Three times weekly

Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vaor phase

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002

Recommended serum: ATCC 30-2020

parental cell line: ATCC CCL-89

0.25% (w/v) Trypsin - 0.53mM EDTA in Hank's BSS (w/o Ca++, Mg ++): ATCC 30-2101

Phosphate-buffered saline: AtT-20/D16v-F2細胞ATCC 30-2200

Cell culture tested DMSO: ATCC 4-X

References: 22306: Gumbiner B, Kelly RB. Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. Cell 28: 51-59, 1982. PubMed: 6279313

22624: Moore HP, et al. Expressing a human proinsulin cDNA in a mouse ACTH-secreting cell. Intracellular storage, proteolytic processing, and secretion on stimulation. Cell 35: 531-538, 1983. PubMed: 6317196

22798: Burgess TL, et al. The exocrine protein trypsinogen is targeted into the secretory granules of an endocrine cell line: studies by gene transfer. J. Cell Biol. 101: 639-645, 1985. PubMed: 2991303

23251: Buonassisi V, et al. Hormone-producing cultures of adrenal and pituitary tumor origin. Proc. Natl. Acad. Sci. USA 48: 1184-1190, 1962. PubMed: 13874682

32927: Stefana B, et al. Leukemia inhibitory factor induces differentiation of pituitary corticotroph function: an immuno-neuroendocrine phenotypic switch. Proc. Natl. Acad. Sci. USA 93: 12502-12506, 1996. PubMed: 8901611