On-feeder culture medium for human ES/ iPS cell

StemSure® on-feeder hPSC Medium

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).

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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.)

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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.

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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.

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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.

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bFGF

It is an essential factor for maintenance culture of human ES / iPS cells.

Y-27632

StemSure® Series

Human ES / iPS cell detection reagent

Human ES / iPS cell removal reagent

CultureSure® Low molecular compound series

For research use or further manufacturing use only. Not for use in diagnostic procedures.

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