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The alpha TC1 clone 6 cells are useful for studying many aspects of islet biology, including glucagon biosynthesis and cytokine sensitivity.

Alpha TC1 clone 6 is a pancreatic alpha cell line. It was 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.

H-2b; H-2d, expressed

glucagon

glucagon

The parental cell line is less differentiated and produces both glucagon and insulin. Two clonal cell lines, alpha TC1 clone 6 and alpha TC1 clone 9 (ATCC CRL-2350), are more differentiated than the parent line and produce only glucagon RefHamaguchi 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. alpha TC1 clone 6 cells exhibit the most differentiated phenotype and express the highest levels of glucagon. RefHamaguchi 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

Volumes used in this protocol are for 75xa0cm² flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
NOTE: Warm all solutions to 37.0°C prior to use

1. Transfer all medium and floating cells from flask to a 50 mL centrifuge tube.
2. Adherent cells are removed using Cell Dissociation Buffer (an enzyme free buffer; Invitrogen, Catalog No. 13150-016) diluted 1:5 with Hanks Balanced Salt Solution. Add 5.0 mL of diluted cell dissociation buffer per 75xa0cm² flask and gently rock flask to bathe the cells at room temperature for 1 to 2 minutes.
3. Allow the flask to remain at room temperature for 1 to 5 additional minutes until cells have detached from the flask.
4. Firmly tap the flask against palm of hand to dislodge cells.
5. Add 10.0 mL of fresh medium per 75xa0cm² 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.
6. Transfer these cells to the centrifuge tube from Step 1. Centrifuge at 125 x gxa0 for 5 to 10 minutes. Remove medium and resuspend pellet in fresh complete medium.
7. Add appropriate aliquots of cell suspension to new culture vessels.
8. Incubate cultures at 37°C.

Medium renewal: Every 2 to 3 days.

Freeze medium: complete growth medium supplemented with an additional 40% heat-inactivated fetal bovine serum and 5% (v/v) DMSO

Temperature: 37°C

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