How to culture MIN6 cells?
You can culture MIN6 cells in DMEM-High Glucose medium, 15% FBS, 1% P/S, 1% Glutamine (2mM), 20 mM HEPES (diluted from 1M HEPES purchased from Sigma) and 50-55 μM beta-mercaptoethanol (1.75-2.0 μl/ 500 ml of medium). Change medium every 2-3 days.
What are MIN6 cells?
The MIN6 cell line was derived from a mouse insulinoma and is one of a few cell lines that display characteristics of pancreatic beta-cells, including insulin secretion in response to glucose and other secretagogues [10], [11].
How does medium change in cell culture?
To media change, warm up fresh culture media (section 5.1) at 37°C in water bath or incubator for at least 30 mins. Carefully pour of the media from the flask into a waste pot containing some disinfectant. Immediately replace the media with 100 ml of fresh pre-warmed culture media and return to CO2 37°C incubator.
What are INS 1E cells?
Rat insulinoma-derived INS-1 cells constitute a widely used beta-cell surrogate. INS-1E cells can be safely cultured and used within passages 40-100 with average insulin contents of 2.30 +/- 0.11 microg/million cells. Glucose-induced insulin secretion was dose-related and similar to rat islet responses.
How does glucose stimulate insulin release?
Increased intracellular glucose then leads to increased production of ATP, and an increase in the ATP/ADP ratio (1); the increased ATP/ADP ratio leads to closing of the potassium channel and depolarization of the cell (2); and cell depolarization opens a calcium channel (3) which leads to insulin secretion (4).
What does insulinoma mean?
Key points. Insulinomas are tumors in your pancreas. They make extra insulin, more than your body can use. Insulinomas can cause hypoglycemia, or low blood sugar.
How do cells change to medium?
Methods for Changing Cell Media
- Warm cell culture medium and HBSS to 37˚C in the water/bead bath;
- Spray them down with ethanol and place them inside a biosafety cabinet (BSC);
- Spray down incubator doors with ethanol and minimize breathing to diminish contamination risk;
How many cm2 is a 6 well plate?
Useful information for various sizes of cell culture dishes and flasks
| Catalog No. | Surface area (cm2) | |
|---|---|---|
| Dishes | ||
| 150mm | 150468;168381 | 145 |
| Culture plates | ||
| 6-well | 140675 | 9.6 |
What stimulates release of insulin?
Insulin secretion is governed by the interaction of nutrients, hormones, and the autonomic nervous system. Glucose, as well as certain other sugars metabolized by islets, stimulates insulin release.
What hormone stimulates insulin?
Although a variety of gut hormones participate in promoting insulin release,108 gastrointestinal polypeptide (GIP) pancreatic glucagon and the glucagon-like peptides (GLP) play a major role in stimulating insulin release.
How is insulinoma diagnosed?
An insulinoma is a rare pancreatic beta-cell tumor that hypersecretes insulin. The main symptom is fasting hypoglycemia. Diagnosis is by a 48- or 72-hour fast with measurement of glucose and insulin levels, followed by endoscopic ultrasound. Treatment is surgery when possible.
What’s the best way to culture MIN6 cells?
Any suggestions are highly welcomed. Join ResearchGate to ask questions, get input, and advance your work. You can culture MIN6 cells in DMEM-High Glucose medium, 15% FBS, 1% P/S, 1% Glutamine (2mM), 20 mM HEPES (diluted from 1M HEPES purchased from Sigma) and 50-55 μM beta-mercaptoethanol (1.75-2.0 μl/ 500 ml of medium).
How often should I Change my min6 culture medium?
Change medium every 2-3 days. Moreover, MIN6 cells are very sensitive to FBS, therefore, it is important to check a few batches of FBS and see whether the cells grow better with a certain batch of FBS. MIN6 culture conditions for ROS analysis? Hello! i’m trying to measure ROS production in MIN6 cells.
Are there any insulin positive cells in MIN6?
Confocal laser scanning microscopy revealed that MIN6 cells included not only insulin positive cells but also insulin and glucagon or somatostin double positive cells. Glucagon, somatostatin and ghrelin were detectable in the culture medium. The present study clearly demonstrated that MIN6 produce pancreatic endocrine cells.
Where did the MIN6 cell line come from?
The MIN6 cell line was derived from a mouse insulinoma and is one of a few cell lines that display characteristics of pancreatic beta-cells, including insulin secretion in response to glucose and other secretagogues [10], [11].
