Switching from feeder layers to feeder free iPSC derrivation on vitronectin with essential media 8, wanted to know if I can culture directly in essential media 8 after transduction till iPSC develop or if I need to use intermediate medias for support - I am using an MBD3 knockdown in conjunction with miRNA reprogramming factors expressed from lentivirus.
Hello Dustin,
We generally are able to get efficient reprogramming in feeder independant manner using Cytotune reprogramming. You can efficiently reprogramm fibroblasts by re-seeding onto vitronectin and either Essentila 8 media or Essential 6 + 100 ng/mL bFGF from day 8 to day 21 to form colonies. Althought this is most efficient with Cytotune reprogramming, it may be ammendable to other technologies. Please see the following page for various protocols that may assist you in using this media/matrxi combination. Best of luck.
https://www.lifetechnologies.com/us/en/home/references/protocols/cell-culture/stem-cell-protocols/ipsc-protocols.html
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