| Usage | Take a six-well plate as an example
1. Thaw the basic medium and supplements in advance, and rewarm the basic medium and accutase digestive juice to room temperature before the experiment.
2. When iPSC/hESC grows to 70-80% confluence, the cells are in good condition under the microscope.
3. Suck and discard the culture medium.
4. Add 1mL of accutase digestion juice (abs47014935) to each well and digest in a 37 °C incubator for 3min.
5. Take 11mL of EB forming medium and mix 22μL of EB forming supplement to obtain EB forming complete medium, and add 2mL of EB to each well of 3 wells of an ultra-low adsorption 6-well plate to form complete medium.
6. When you see that the clones are white and bright, gently pat the side of the plate (twice on each side), pat the cells down, and then add 1mL of EB to form a complete medium in the biological safety cabinet to terminate digestion. Suck this 2mL system into a 15mL centrifuge tube filled with 3mL of EB to form a complete medium and centrifuge 160g for 5min.2Overnight.
7. On the next day, it can be seen that a large number of uniform EB is formed under the microscope. Half of the solution is changed, 50% of the EB formation medium (excluding supplements) is supplemented, and it is placed on a horizontal shaker for culture.
8. Change the basal medium every day, or conduct subsequent differentiation experiments. |
| Description | Embryoid body differentiation kit, capable of supporting human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) Generate embryoid bodies (EB) and ensure their survival in the subsequent culture process, providing support for subsequent differentiation experiments.
The kit does not contain serum components, and has been verified many times by H1, H9 and iPSC cells, and can stably form embryomimetic bodies in large quantities.
Product composition:
| serial number | Name | Specifications | Save | | Component A | EB basal medium | 200mL | -20℃ | | Component B | EB Supplement (500 ×) | 400uL | -20℃ |
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| General Notes | 1. The optimum density of iPSC/hESC to form embryoid bodies is about 1.5 * 105Cells/mL, for six-well plates, when one well of cells grows to 70-80% confluence, it can be used for embryomimetic body formation in three wells of six-well plates.
2. Formation of embryoid iPSC/hESC cells remain undifferentiated. Please make sure that the cells are in good condition before the experiment.
3. A large number of embryoid bodies can be seen on the second day, but the individuals are small, so it is not recommended to conduct experiments such as differentiation directly. It is recommended to conduct experiments on the third day. |
| Storage Temp. | Stored at-20 ℃, valid for 6 months; Thaw at 2 ~ 8 ℃ and store for 2 weeks. Repeated freezing and thawing are prohibited. |
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