StemSure® on-feeder hPSC medium is a liquid medium used for culturing human ES/ iPS cells on feeder cells. Little preparation is needed as it is a pre-mixed medium that contains the basal medium, serum replacements, non-essential amino acids, and reducing agents. bFGF should be added prior to use as it is not contained in the mixture.
Feature
- Pre-mixed medium, ready-to-use with the addition of bFGF
Validation Tests
- Appearance
- pH
- Osmotic pressure
- Sterility test
- Endotoxin test
- Mycoplasma test
- Cell proliferation assay
- Alkaline phosphatase staining
Colony Formation and ALP Staining 1
Human iPS cell line 201B7 maintained in a feeder-free medium was transferred to be cultured on feeder cell layer in either StemSureStemSure on-feeder hPSC medium or an equivalent product from another company. Morphology of the colonies and alkaline phosphatase (ALP) staining pattern were examined after 5 passages (see below). ALP staining demonstrated that there were no colonies with differentiated cells even after 5 passages.
There were no abnormal findings in terms of the morphology of colonies for cells grow in StemSureStemSure on-feeder hPSC medium after 15 passages (final concentration of bFGF: 5 ng/mL, coating: gelatin).
Colony Formation and ALP Staining 2
Human iPS cell line 201B7 was cultured for 19 passages in either StemSureStemSure on-feeder hPSC medium or an equivalent product from another company. Using our product, undifferentiated state of the cells was maintained for a prolonged period of time.
(※ % indicated in the right bottom corner of ALP images indicate the % of differentiated colonies.)
Cell Proliferation
Human iPS cell line 201B7 was cultured in StemSureStemSure on-feeder hPSC medium. Cell proliferation was observed for 3 days using IncuCyte ZOOM.
Identifying expression of markers for undifferentiated cells
Human iPS cell line 201B7 maintained in a feeder-free medium was transferred to be cultured on feeder cell layer in StemSureStemSure on-feeder hPSC medium. After 5-6 passages, cells were stained with markers of undifferentiated cells (OCT3/4, SOX2, TRA-1-81, and BC2LCN) to confirm that undifferentiated states were maintained.
※BC2LCN is a recombinant lectin that is highly specific to sugar chains on the surface of undifferentiated human ES/ iPS cells.
Confirming differentiation into three germ lineages
Human iPS cell line 201B7 was cultured in StemSureStemSure on-feeder hPSC medium for 9 passages, and were seeded on a gelatin-coated dish 7 days after the formation of EB. Cells were cultured for 14 days, and the expressions of βⅢ-Tublin, α-SMA, and AFP were examined to confirm differentiation into three germ lineages.
