I'm expading MSC (Mesenchymal Stem Cell) from umbilical cord uner 37 °C with 5% CO2 in a humidified chamber. I'm testing 2 types of media, both appropriate for MSC (StemPro and NutriStem). I need to expand the cells, but they keep differentiating. Does anyone know what might be happening?
Dear Leticia,
What do you mean by differentiating? Did you checked using alcian blue staining for chondrogenic, alizarin red staining for osteogenic or oil red O staining for adipogenic differentiation? I supposed that what you meant was that your cells did not grow in monolayer, but tended to form clumps of several layers, was that so?. Stem Pro or other comercial serum free medium do not contain attachment factor, so you need to coat your flask with collagen or fibronectin, to allow your cells attach to the flask and grow nicely as monolayer. Different if you use serum or platelet lysate containing medium. Serum and platelet lysate contain attachment factor, so you do not need to coat the flask, but the cells will grow nicely as a monolayer
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