INS-1 Cells
Product number:
300471
General information
Description | The INS-1 cell line is derived from an x-ray-created transplantable insulinoma in rats. Because INS-1 cells contain a high concentration of insulin and respond to changes in glucose levels, they are frequently used to study the function of beta cells. Growth and hormone expression are dependent on the reducing agent 2-mercaptoethanol. INS-1 cells are notable for their heterogeneity, consisting of mature insulin-positive cells and immature bi-hormonal cells expressing insulin and glucagon proteins. Bi-hormonal INS-1 cells have lower Nkx6.1 expression and lack alpha cell markers, indicating they are not fully matured. Furthermore, chronic glucose stimulation reduces insulin gene and protein expression in INS-1 cells. As such, insulin and proglucagon-derived peptides like GLP-1, GLP-2, and glucagon levels are reduced. |
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Organism | Rat |
Tissue | Pancreas, islets of Langerhans |
Disease | Rat insulinoma |
Synonyms | INS1 |
Characteristics
Age | 666 days |
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Gender | Male |
Cell type | Beta cell |
Growth properties | Adherent/suspension |
Identifiers / Biosafety / Citation
Citation | INS-1 (Cytion catalog number 300471) |
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Biosafety level | 1 |
Expression / Mutation
Products | Insulin, glutathione |
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Handling
Culture Medium | RPMI 1640, w: 4.5 g/L Glucose, w: 2 mM L-Glutamine, w: 10 mM HEPES, w: 1 mM Sodium pyruvate, w: 1.5 g/L NaHCO3 (Cytion article number 820702a) |
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Medium supplements | Supplement the medium with 10% heat-inactivated FBS |
Passaging solution | Accutase |
Subculturing | Gather the suspension cells in a 15 ml tube and gently wash the adherent cells with PBS lacking calcium and magnesium (use 3-5 ml for T25 flasks and 5-10 ml for T75 flasks). Apply Accutase (1-2 ml for T25 flasks, 2.5 ml for T75 flasks) ensuring full coverage of the cell layer. Allow the cells to incubate at room temperature for 10 minutes. Following incubation, combine and centrifuge both the suspension and adherent cells. After centrifugation, carefully resuspend the cell pellet and transfer the cell suspension into new flasks containing fresh medium. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures | INS-1 cells are shipped in a deep-frozen state on dry ice. Upon receipt, confirm that the vial remains frozen. For storage, place the cryovial immediately at temperatures below -150 degrees. If you plan to culture the cells immediately, swiftly thaw the vial by shaking it in a 37 degrees water bath with clean water and an antimicrobial agent for 40-60 seconds. Remove the vial once a small ice clump persists, ensuring it remains cold. Proceed with all subsequent steps under aseptic conditions. In a sterile flow hood, disinfect the cryovial with 70% ethanol. Then, gently open the vial and transfer the cell suspension into a 15 ml centrifuge tube pre-filled with 8 ml of room temperature culture medium. Gently mix the cells. For cell separation, centrifuge at 300 x g for 3 minutes and dispose of the supernatant. Skipping centrifugation is optional, although any residual freezing medium should be removed after 24 hours. Resuspend the pellet gently in 10 ml of fresh culture medium and divide between two T25 culture flasks. Follow the subculture protocol for subsequent steps. |
Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is rigorously excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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