HybriS-8 medium with alanyl-glutamine and supplement, 490 + 10 mL, EuroPET + PET
A mixture of mineral salts, amino acids, glucose and vitamins dissolved in deionized water and sterilized by membrane filtration. HybriS-8 contains no antibiotics, albumin, fetuin or components of undefined composition, such as plant or animal protein hydrolysates.
Appearance: A clear, reddish-orange liquid without opalescence or precipitate.
Intended use: Serum-free, feeder-free culture of induced pluripotent stem cells (iPSCs) and embryonic stem cells (ES cells). Supports proliferation and maintenance of pluripotency over extended passage series. Used in cell biology, genetics and regenerative medicine research.
Directions for use
- Open the medium bottle under aseptic conditions.
- Add HybriS-8 supplement (50×) at a dilution of 1:50.
- Add antibiotics if required.
Stem cells cultured in HybriS-8 can be passaged without exposure to serum using trypsin and soybean trypsin inhibitor. An enzyme-free protocol based on Versene treatment can also be used.
For long-term routine culture, replace the medium daily. Use an appropriate commercially available protein coating for cell culture plasticware. Passage cells as small aggregates (50–200 µm) when colonies cover 60–70% of the available surface or when very large colonies with multilayered centres develop. Dilute the cell suspension from 1:3 to 1:12, depending on the required seeding density and culture schedule. Single-cell passaging with a ROCK inhibitor (Y-27632) is also possible, but may reduce viability and pluripotency.
Storage: Store the medium at +2 to +8°C, protected from light. Store the supplement frozen at −15 to −20°C. Transport by any covered means of transport.
HybriS-8 performance in a model experiment
In a model experiment, HybriS-8 supported iPSC growth for at least 8 passages. Within 5–7 days of seeding, colonies increased 10–50-fold without signs of differentiation. Proliferation was accompanied by maintenance of morphology and a high growth index (GI = 10.1).
This product is intended for research use.
