How to use Gibco PSC Neural Induction Medium
3:14
With Gibco PSC Neural Induction Medium, you can efficiently convert 1 million human pluripotent stem cells, or PSCs, into 20 million neural stem cells without laborious embryoid body formation or rosette isolation. Before neural induction, the quality of human PSCs is critical for highly efficient neural induction. High-quality PSCs with minimal or no differentiated colonies, as shown in the images, grown in feeder-free media, such as Essential 8 Medium or StemFlex Medium, are recommended. Any differentiated or partly differentiated colonies, as pointed with the arrow, should be removed before passaging. When PSCs reach 70% to 80% confluence,as shown in the image, passage and plate PSCs at a density of 25,000 to 30,000 cells/cm2 in a coated tissue culture dish. The use of 10 µM ROCK inhibitor during passaging has been shown to improve cell viability and attachment at this step. 24 hours after PSC splitting, the density of PSCs should be 15% to 25% confluence, as shown in the image. Simply switch the growth medium to Gibco PSC Neural Induction Medium to start neural induction. The total duration of neural induction is 7 days. Change Neural Induction Medium every other day during the 7 days of neural induction. During the 7 days, colonies with a uniform morphology indicate good neural induction. During neural induction, cells are keeping to proliferate, and cell density are increasing during induction process. As shown in the images, colonies with a non-neural morphology may be observed during neural induction due to differentiated PSCs being in the culture before neural induction. To remove them, mark the differentiated colonies with a microscopy marker and aspirate them with a Pasteur glass pipette when changing the medium. At day 7 of neural induction, cells reach confluence, as shown in the images. Induced NSCs can then be disassociated and expanded in neural expansion medium. Expanded NSCs can be stained with antibodies against neural markers, such as Sox1, nestin, and Sox2. Expanded NSCs can also be differentiated into specific neural cell types, such as neurons, astrocytes, and oligodendrocytes. Gibco Neural Induction Medium generates at least 20 million NSCs from 1 million PSCs in 7 days.
