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Add 1/1,000 volume of this product to the culture medium and mix thoroughly before use.
Example: 10 mL of hPSCs medium + 10 μL of this product
Precautions for Use
Samples: human ES cells, human iPS cells
Applications:
The human iPS cell 201B7 strain was seeded in StemSure hPSC Medium Δ containing bFGF supplemented with CEPT or Y-27632 at 37°C under 5% CO2 conditions (6-well plates, 1 well= 9.5 cm2).
After at least 16 hours from the start of culture, the medium was replaced with the medium without CEPT or Y-27632, and the culture was continued with media changes every 2 to 3 days.
After checking the cells on day 8, they were fixed with 4% paraformaldehyde, stained with ALP, and colonies were counted.
The human iPS cell 201B7 strain was seeded in StemSure hPSC Medium Δ containing bFGF supplemented with CEPT or Y-27632 at 37°C under 5% CO2 conditions (96-well plates).
After at least 16 hours from the start of culture, the medium was replaced with the medium without CEPT or Y-27632, and the culture was continued with media changes every 2 to 3 days.
After checking the cells on day 13, they were fixed with 4% paraformaldehyde, stained with ALP, and the number of wells with and without colonies were counted. The colony formation rate was calculated from the number of wells with colonies relative to the total number of wells.
Result: When the number of cells seeded was small, more colonies were formed in the CEPT-supplemented cultures than in the Y-27632-supplemented cultures.
Cells: human iPS cell 201B7 strain
Medium: StemSure hPSC Medium Δ + 35 ng/mL bFGF
Red: rBC2LCN-635 (human iPSC membrane stain)
Blue: DAPI (nuclear stain)
Note: BC2LCN is a recombinant lectin with high affinity for cell surface glycans of human ES cells and human iPS cells.
Result: No difference in cell morphology was observed between the addition of CEPT and Y-27632. The undifferentiated state was also maintained.
▽ With CEPT
▽ With Y-27632