| 產品名稱 |
alphaTC1 Clone 9 |
| 商品貨號 |
B163932 |
| 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)]
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 cm 2 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
transgenic for the SV40 T antigen
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