Amniokar Medium
It is intended for the cultivation of amniotic fluid cells and chorionic cell elements in vitro using "In situ" or "Flask" methods in order to obtain an enriched population of dividing cells and subsequent analysis of metaphase chromosomes. It is highly effective at the cultivation of fibroblasts, mesenchymal stem cells (MSCs) and other cells of mesenchymal origin.
Stopping mitotic division with subsequent accumulation of metaphase plates is achieved by incubation of cells with colchicine. For analysis, the cells are transferred to hypotonic solution of potassium chloride, fixed, applied to a slide and stained, according to generally accepted methods.
This product is intended for research purposes. The product has a Russian registration certificate as a medical product.
Appearance: clear, red-orange solution, without opalescence and sediment.
"Flask" cultivation method:
- Before use, carefully mix the contents of the bottle with the Amniokar medium (especially in the case of using a refrozen product!).
- 10 ml of freshly obtained (storage no more than 4 - 8 hours at a temperature from 30 to 37 °C) amniotic fluid is centrifuged at 1000x g for 10 minutes.
- Select and remove the supernatant (leaving ~1-2 ml of supernatant).
- Carefully resuspendthe pellet, add 5 ml of Amniokar solution, mix gently and transfer the entire volume of liquid into a ventilated culture vial, with 25 cm² bottom area.
- Cell cultivation is carried out in a CO₂ incubator, at temperature 37 ±0.5 °C, CO₂ concentration - 5%.
- On the 5th day, add 50% volume of fresh Amniokar Medium to the culture.
The total incubation time is 9-12 days from the culture passage start.
- Mitotic cell division is stopped by treating cell culture with Colchicine (concentration - 1 mcg/ml, incubation for 60 minutes).
- Removal of cells from the plastic surface is carried out according to the standard protocol.
- For analysis, the obtained cell suspensions are centrifugated at 1000-1500x g for 10 min. Then the cells are treated with hypotonic solution, fixed, preparations are prepared and stained according to the standard protocols.
HOW SUPPLIED: in sterile plastic bottles of 60, 100 or 500 ml. Euroflasks have a wide neck, convenient for work.
SHELF LIFE: 1 year. Shelf life after opening: 60 days.
STORAGE CONDITIONS: -10 to -20 °C in a place protected from light. After opening store at temperature +2 to +8 °C.
RECOMMENDED USE:
- Do not use the product after its expiry date.
- Use gloves, mask and clothes adapted to the manipulation of the product to avoid contamination.
- Manipulate the product in aseptic conditions.
- Medium should be clear and free of opalescence and sediment.
