alphaTC1 Clone 9 (ATCC® CRL-2350)

Organism: Mus musculus, mouse  /  Cell Type: alpha cell  /  Tissue: pancreas  /  Disease: adenoma

Permits and Restrictions

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Organism Mus musculus, mouse
Tissue pancreas
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)]
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 (Invitrogen Cat. No. 31600-034). To make the complete growth medium, add the following components to the base medium:
  • heat-inactivated fetal bovine serum (FBS) to a final concentration of 10%
  • 15 mM HEPES
  • 0.1 mM non-essential amino acids
  • 0.02% bovine serum albumin
  • 1.5 g/L sodium bicarbonate
  • 2.0 g/L glucose

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.

  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). 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.
  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 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.
  6. 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.
  7. Add appropriate aliquots of cell suspension to new culture vessels.
  8. 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
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Name of Depositor EH Leiter
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

transgenic for the SV40 T antigen

Basic Documentation
FAQ's
  1. CRL-2350 growth and morphology
    CRL-2350 is a difficult cell line to grow.  These cells grow as poorly attached single cells and clusters which may also become ...
    Date Updated: 2/21/2014
Restrictions

These cells are distributed for research purposes only. 1. The alphaTC1 Clone 9 cell line was deposited for research purposes only. Neither the cell line nor products derived from it may be sold or used for commercial purposes. Nor can the cells be distributed to third parties for purpose of sale, or producing for sale, cells or their products. The cells are provided as a service to the research community. They are provided without warranty or merchantability of fitness for a particular purpose or other warranty, express or implied. 2. Any proposed commercial use of these cells or products produced by them must first be negotiated with Edward H. Leiter, Telephone: 207-288-6370, FAX: 207-288-6077. 3. In all papers reporting any use of these cells or derived products, a direct reference will be made to the publications below.

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

transgenic for the SV40 T antigen