One of the most remarkable approaches of stem cell culture is xeno-free stem cell culture, a technique that fosters unprecedented growth and differentiation of stem cells while eliminating the use of animal-derived components. Creative Biolabs shares this comprehensive protocol that includes both experiment steps and operating tips, guaranteeing enlightenment in xeno-free culture.
Combine DMEM/F12 with HSA or HS. Add recombinant human proteins, such as bFGF, EGF, and TGF-β. Supplement the medium with ITS, L-ascorbic acid 2-phosphate, and lipids. Filter sterilize the medium, and store the medium at 4°C in the dark for up to two weeks.
Thaw the required amount of laminin or fibronectin at 4°C. Dilute the protein in a sterile buffer (e.g., PBS) according to the manufacturer's instructions. Add the diluted protein to culture dishes or plates and incubate at 37°C. Aspirate the excess protein solution before seeding the stem cells.
Gently dissociate the stem cells using enzymatic or non-enzymatic methods. Count the cells and adjust the concentration to the desired density in xeno-free culture medium. Seed the cells onto the pre-coated xeno-free culture dishes or plates. Place the dishes or plates in a 37°C incubator with 5% CO2. Change the medium daily or every other day, depending on cell density and growth rate.
By implementing this protocol, we wish you are paving the way for groundbreaking research and therapeutic applications. Creative Biolabs stands beside you, offering unparalleled support in your pursuit of advancements in the field of stem cell biology. Contact us for more technical support.
References
For Research Use Only. Not For Clinical Use.
