Feeder free medium for ES/iPS cells StemFit

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StemFit™ Basic03

StemFit™ Basic03

StemFit Basic03 is an animal-origin free, defined medium for human pluripotent stem cell culture. This medium is capable of maintaining human ES and iPS cell under feeder-free culture conditions.

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Animal-Origin Free & regulatroy compliance
for CGT product manufacturing

Animal or human derived components, such as serum derived albumin, or animal cell derived recombinant proteins, are known to carry a risk of hazardous viral contamination for cell therapy. Therefore, StemFit is designed and manufactured under a strict animal origin free policy and is free from animal or human derived components, as referred in USP<1043> and ISO20399. This makes StemFit Basic03 a gold-standard hPSC expansion media for clinical research. To this end, the Japanese PMDA was consulted on the formulation and manufacturing process and found that Basic03 satisfies all requirement of ancillary materials for cell and gene therapy (CGT) manufacturing. Additionally, StemFit offers a GMP compliant product, Basic03 GMP, which is manufactured under applicable GMP guidelines and is thus preferable for CGT products.

Superior performance in single-cell expansion

Single-cell expansion is an ideal method for CGT product manufacturing since it allows well-controlled, reproducible, and efficient cell expansion. StemFit is optimized for the single-cell expansion method and shows higher cell survival rate in single-cell conditions, which enables efficient and genetically stable cell growth without karyotypic abnormalities.
CGT Catapult, an independent center of excellence to advance the growth of the UK CGT industry, performed a comprehensive comparison program for hPSC culture media and found that StemFit Basic03 to have the highest performance in clinical hPSC expansion.

Figure 1. Easy expansion

Figure 2. Consistent gene expression profile

Table 2. Result of karyotyping (CGH array) analysis after expansion

Weekend-Free Feeding

Traditional culture medium requires daily medium changes to maintain hPSCs, which increases the labor cost and risk of human error or contamination in CGT product manufacturing. StemFit allows flexible weekend-free feeding in which hPSC can be expanded healthily without medium change during the weekends. This feature enables researchers to simplify their culture protocol and minimize the labor and culture media costs.

Skip feeding on weekends for flexible scheduling

— Materials Provided —1

Component Volume Storage
Liquid A 400ml Store at below -20 °C
Liquid B 100ml Store at below -20 °C

— Media Preparation —

StemFit Basic03 is provided frozen as a 2 component set containing “Liquid A” and “Liquid B” and can be stored at below -20 °C until use. Use sterile techniques to prepare StemFit Basic03 medium.

1) Before use, thaw the frozen “Liquid A” and “Liquid B” with occasional mixing at room temperature (15-25 °C).
Do not thaw, “Liquid B” at 37 °C, as it accelerates the degradation ingredients. 2

2) Aseptically mix medium components by adding the full volume of “Liquid B” to “Liquid A”. Mix thoroughly.
If prepared aseptically, StemFit Basic03 is ready for use. If precipitations are observed, keep the bottle at room temperature until completely dissolved.

3) Add bFGF at 80 ng/ml concentration*.

4) Upon thawing, StemFit Basic03 medium may be aseptically aliquoted and stored at below -20 °C.

5) Before use, warm aliquots to room temperature and use immediately.

6) nce aliquots are thawed, they should be stored at 4 °C for no more than 2 weeks.34
*bFGF concentration can be customized according to the application.

Key points for successful culture

bFGF not contained

This product does not contain basic fibroblast growth factor
(bFGF). Therefore, 80 ng/ml bFGF should be supplemented.

not 37°C

Do not thaw, “Liquid B” at 37 °C, as it accelerates the degradation of medium ingredients.

2weeks

Thawed StemFit Basic03 medium may be stored at 2-8 °C for up to two weeks.

not Light

We recommend storing the medium protected from light

— Precaution and Disclaimer —

StemFit Basic03 is for research use only and is not for diagnostic use, therapeutic use, or as a food.

For Research Use Only. Not Intended For Human or Animal Diagnostic or Therapeutic Uses
Download

・Product Information (PDF [346KB])

・SDS (PDF [362KB] )

・Instruction Manual
StemFit medium (PDF [775KB] )
StemFit medium with iMatrix-511 (PDF [2.4MB] )

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StemFit™ Basic03
FAQ

  • Which bFGF can be used for expansion? Ajinomoto provides high quality, animal-origin free bFGF, however commercially available bFGF have been confirmed to work.
  • Which Extracellular Matrix (ECM) is recommended? Any commercially available ECMs for hPSCs will work (e.g. iMatrix-511, Geltrex, VTN-N, LN521, Matrigel and Synthemax) .
  • Can I use Basic03 for clump passaging? We recommend single-cell passaging.  If you do passage cells in clumps, we recommend to make small clumps and seed at low seeding density.
  • How should I switch medium to Basic03? We recommend to switch to Basic03 2 days before passaging.
    Please refer to Technical Tips for more details
  • Can I use Basic03 for non-human PSCs? StemFit medium has been developed for human PSCs, however there are reports in which primate PSCs have been cultured in StemFit medium
  • Can I use Basic03 for Mesenchymal stem cell or hematopoietic stem cells? No. StemFit For MSC medium is available to expand MSC.
  • Is GMP medium available? Basic03 GMP medium is available. Basic03 GMP correspond to Basic03 and it is manufactured under GMP guidelines.
  • How many passages can Basic03 be used to maintain PSCs? We have confirmed that PSCs cultured in Basic03 maintain their normal karyotype after 20 passages.
  • What’s the difference between Basic03 and AK03N? AK03N is available only in Japan and corresponds to Basic03 but it includes bFGF. Basic03 is used for iPSC establishment, expansion and is also used in a differentiation process as a basal media.
Differentiation medium MSC medium Recombinant Proteins

For more information, please contact us.

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AJINOMOTO CO., INC. AminoScience Division
15-1, Kyobashi 1-Chome, Chuo-Ku, Tokyo 104-8315, Japan

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