HiDef® S8 stem cell suspension medium

Product background

HiDef® S8 is a unique formulation that enables high-yield, scalable suspension expansion of human pluripotent stem cells without the need for microcarriers. The medium supports efficient aggregate formation from single-cell or small aggregate inoculation, rapid biomass accumulation, and expansion to tens of millions of PSCs per milliliter while maintaining high viability, uniform aggregate morphology, stable size distribution, and expression of key pluripotency markers. Following suspension expansion, cells readily transition back to adherent culture while preserving normal morphology and pluripotent characteristics.

HiDef® S8 contains a combination of essential nutrients and growth factors formulated specifically to maintain pluripotency while supporting robust suspension growth under agitation conditions. The addition of further components is not required, although optional surfactants such as polyvinyl alcohol (PVA), Pluronic® F-68, dextran sulfate, and others may be incorporated where desired for specific suspension workflows.

HiDef® S8 400x Supplement has been verified for compatibility with standard formulations of DMEM/F12 from several manufacturers, related additives may be used depending on the application and bioreactor configuration.

Each lot of HiDef® S8 400x Supplement is used in combination with basal medium during performance qualification using multiple human PSC lines cultured under suspension conditions in both ambient and hypoxic environments.

Key characteristics

HiDef® S8 complete medium is a specially formulated defined medium that supports feeder-free, serum-free expansion of human pluripotent stem cells in high-density suspension culture while enabling scalable manufacturing workflows.

• Defined, serum-free, feeder-free medium for high-density human PSC suspension culture
• Supports microcarrier-free expansion in stirred bioreactors and other suspension systems
• Enables rapid biomass generation while maintaining high viability and pluripotency
• Compatible with single-cell inoculation and Ready-CEPT® supplementation
• Supports efficient transition back to adherent culture following suspension expansion
• Scalable from milliliter-scale research workflows to multi-liter bioreactor production

Preparation of complete HiDef® S8 medium (standard vial)

Use appropriate aseptic technique to prepare HiDef® S8 complete medium (DMEM/F12 basal medium + HiDef® S8 400x supplement). The following example is for preparing 501.25 ml of HiDef® S8 complete medium. If preparing other volumes, adjust accordingly.

NOTE: Thaw supplements at room temperature (15–25°C) and use immediately. Do not thaw in a 37°C water bath, overnight at 4°C, or on ice.

1. Thaw the HiDef® S8 400x Supplement and allow it to warm to room temperature. Mix thoroughly by gentle inversion or pipetting. Turbidity may be observed after thawing; this is normal and should fully resolve upon dilution in basal medium (Step 2).
2. Add 1.25 ml of HiDef® S8 400x Supplement (the full vial volume) to 500 ml of DMEM/F12. Mix thoroughly while avoiding foaming.
3. When prepared aseptically, HiDef® S8 complete medium is ready for immediate use. Store at 2-8°C for up to two weeks. Do not freeze or refreeze prepared HiDef® S8 complete medium.

Preparation of complete HiDef® S8 medium (miniature vial)

Defined Bioscience also offers HiDef® S8 in a miniature format, enabling convenient preparation of small volumes for pilot studies, optimization experiments, and low-throughput suspension culture workflows. Each miniature 400x Supplement vial prepares 50.125 ml of complete HiDef® S8 medium. Miniature 10-packs collectively yield 501.25 ml of complete medium.

1. Thaw the HiDef® S8 400x supplement mini and allow it to warm to room temperature. Mix thoroughly by gentle inversion or pipetting. Turbidity may be observed after thawing; this is normal and should fully resolve upon dilution in basal medium (Step 2).
2. Add 0.125 ml of HiDef® S8 400x supplement mini (the full vial volume) to 50 ml of DMEM/F12 in a sterile 50 ml conical tube or equivalent vessel. Mix thoroughly while avoiding foaming.
3. When prepared aseptically, HiDef® S8 complete medium is ready for immediate use. Store at 2-8°C for up to two weeks. Do not freeze or refreeze prepared HiDef® S8 complete medium.