alphaTC1 Clone 9
CRL-2350 ™
alphaTC1 Clone 9 is an alpha cell that was isolated from the pancreas of a mouse with adenoma. This cell line can be used for studying glucagon biosynthesis and alpha cell sensitivity to cytokines.
- Product category
-
Animal cells
- Organism
-
Mus musculus, mouse
- Classification
- Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
- Cell type
- alpha cell
- Morphology
- epithelial
- Tissue
- Pancreas
- Disease
-
Adenoma
- Applications
-
3D cell culture
- Product format
- Frozen
- Storage conditions
-
Vapor phase of liquid nitrogen
Documentation
ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
Cells contain SV40
ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
Detailed product information
General
- Specific applications
-
This cell line is useful for studying glucagon biosynthesis and alpha cell sensitivity to cytokines.
Characteristics
- Growth properties
- Adherent, single cells and loosely attached clusters
- 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)].
- Strain
-
(C57BL/6J x DBA/2)F2
- Antigen expression
-
H-2bH-2d
- Genes expressed
- 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.
Handling information
- Unpacking and storage instructions
-
- Check all containers for leakage or breakage.
- Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below -130°C, preferably in liquid nitrogen vapor, until ready for use.
- Complete 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%
- Temperature
-
37°C
- Atmosphere
-
90% Air, 10% CO2
- Handling procedure
-
To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
- Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
- Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
- It is recommended that the cryoprotective agent be removed immediately. Centrifuge the cell suspension at approximately 125 x g for 5 to 10 minutes. Discard the supernatant and resuspend the cell pellet in an appropriate amount of fresh growth medium. to one T-25. Incubate the culture at 37°C in a suitable incubator. A 10% CO2 in air atmosphere is recommended if using the medium described on this product sheet.
- Subculturing procedure
-
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. - Reagents for cryopreservation
- Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)
Quality control specifications
- Mycoplasma contamination
- Not detected
History
- Deposited as
- Mus musculus
- Depositors
- EH Leiter
Legal disclaimers
- Intended use
- This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
- Warranty
-
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
- Disclaimers
-
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.
While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.
This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.
Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.
Permits & Restrictions
Import Permit for the State of Hawaii
If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.
Frequently Asked Questions
References
Curated Citations
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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