The first xeno-free supplement for hESC and hiPSC culture

Gibco™ CTS™ KnockOut™ SR XenoFree Medium is a xeno-free culture supplement that maintains the pluripotency, normal morphology, and karyotype of human PSCs (Figure 1). It exhibits performance equivalent to that of traditional Gibco™ KnockOut™ Serum Replacement and supports the derivation and routine maintenance of human ESCs and iPSCs.

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CTS KnockOut SR XenoFree supports:

  • Feeder-dependent culture of PSCs with human feeder cells and Gibco™ CELLstart™ substrate
  • Additional applications such as cryopreservation, derivation, and differentiation
  • Reduced burden of qualifying reagents when transitioning to clinical applications

Figure 1. hESCs cultured in KnockOut SR XenoFree Medium retain their pluripotent phenotype. BG01v cells cultured using CTS KnockOut SR XenoFree Medium maintain expression of common hESC markers. BG01v cells were cultured on human foreskin fibroblasts and CTS™ CELLstart™ Substrate for 5 passages prior to immunocytochemical staining. (A) Phase contrast; (B) DAPI; (C) Oct4; (D) SSEA4 Alexa Fluor™ 488 and 555, respectively.

Figure 2. Xeno-free culture and gene expression of hESCs. (A) Xeno-free culture of hESCs on feeder cells. BG01v morphology when cultured using CTS KnockOut SR XenoFree Medium on human foreskin fibroblasts (HFF) attached with CTS CELLstart Substrate, passage 4. (B) Maintenance of pluripotency with CTS KnockOut SR XenoFree Medium. Following 10 passages in either medium supplemented with KnockOut Serum replacement (left lane of the pair for each marker) or CTS KnockOut SR XenoFree medium (right lane of the pair for each marker) on HFF attached with CTS CELLstart Substrate, BG01v gene expression was examined (top). Gene expression of embryoid bodies generated from the same passage-10 BG01v/HFF culture (bottom).

Complete media system

Supplement Basal medium Recommended reagent
CTS KnockOut SR XenoFree Medium CTS KnockOut DMEM CTS CELLstart substrate