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alphaTC1 Clone 9

規(guī)格:

價格:

編號:B163932

品牌:Mingzhoubio

標準菌株

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

alphaTC1 Clone 9

商品貨號

B163932

Organism

Mus musculus, mouse

Tissue

pancreas

培養(yǎng)基

Cell Type

alpha cell

Product Format

frozen

Morphology

epithelial

Culture Properties

adherent, single cells and loosely attached clusters

Biosafety Level

2 [Cells Contain PAPOVAVIRUS (SV-40)]

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.

Disease

adenoma

Strain

(C57BL/6J x DBA/2)F2 (transgenic for the SV40 T antigen)

Applications

This cell line is useful for studying glucagon biosynthesis and alpha cell sensitivity to cytokines.

Storage Conditions

liquid nitrogen vapor phase

Derivation

alphaTC1 clone 9 is a pancreatic alpha cell line cloned from the alpha TC1 cell line which was derived from an adenoma created in transgenic mice expressing the SV40 large T antigen oncogene under the control of the rat preproglucagon promoter.

This cell line is heterozygous for MHC alleles derived from both parental strains used in construction of the transgenic mice [C57BL/6J(H-2b] and DBA/2J(H-2d)].

Antigen Expression

H-2b

H-2d

Genes Expressed

glucagon

Cellular Products

glucagon

Comments

Though the parental alphaTC1 cell line produces glucagon and considerable quantities of insulin and preproinsulin mRNA, the clonal line is more differentiated and produces glucagon but not insulin or preproinsulin mRNA.

Rat recombinant gamma-interferon or mouse recombinant interleukin-1 individually inhibited glucagon synthesis in alphaTC1 clone 9 cells but did not cause inhibition of DNA synthesis.

The combination of these cytokines caused marked inhibition of DNA and glucagon synthesis in alphaTC1 clone 9 cells.

Complete Growth Medium

The base medium for this cell line is Dulbecco's Modified Eagle's Medium, low glucose (Gibco Cat. No. 11885-084). To make the complete growth medium, add the following components to the base medium:

Fetal bovine serum (FBS) to a final concentration of 10%
HEPES to a final concentration of 15 mM
Non-essential amino acids to a final concentration of 0.1 mM
Bovine serum albumin to a final concentration of 0.02%

Subculturing

Volumes used in this protocol are for 75 cm² flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
NOTE: Warm all solutions to 37°C prior to use.

Transfer all medium and floating cells from flask to a 50 mL centrifuge tube.
Adherent cells are removed using Cell Dissociation Buffer (an enzyme free buffer; Invitrogen, Catalog No. 13150-016). Add 5 mL of diluted cell dissociation buffer per 75 cm² flask and gently rock flask to bathe the cells at room temperature for 1 to 2 minutes.
Allow the flask to remain at room temperature for 1 to 5 additional minutes until cells have detached from the flask.
Firmly tap the flask against palm of hand to dislodge cells.
Add 10 mL of fresh medium per 75 cm² flask and triturate up and down directing the stream along the growth surface of the flask to dislodge the cells and break up some of the clumps.
Transfer these cells to the centrifuge tube from Step 1. Centrifuge at 125 x g for 5 to 10 minutes. Remove medium and resuspend pellet in fresh complete medium.
Add appropriate aliquots of 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: 2 to 3 times a week.

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.

Cryopreservation

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

Storage temperature: liquid nitrogen vapor phase

Culture Conditions

Temperature: 37°C

Atmosphere: air, 90%; carbon dioxide (CO₂), 10%

Name of Depositor

EH Leiter

Deposited As

Mus musculus

References

Powers AC, et al. Proglucagon processing similar to normal islets in pancreatic alpha-like cell line derived from transgenic mouse tumor. Diabetes 39: 406-414, 1990. PubMed: 2156740

Hamaguchi K, Leiter EH. Comparison of cytokine effects on mouse pancreatic alpha-cell and beta-cell lines. Viability, secretory function, and MHC antigen expression. Diabetes 39: 415-425, 1990. PubMed: 2108069

transgenic for the SV40 T antigen

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