StemFit® Feeder-Free Stem Cell Culture Media
Allowing weekend-free culture of stem cells
StemFit® is a range of animal-origin free, chemically defined media proven to effectively maintain induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) under feeder-free conditions during the reprogramming, expansion and differentiation phases of stem cell culture.
Available in research and clinical grade formulations, StemFit® Stem Cell Culture Media combines market-leading colony forming efficiency with lower than standard media volume consumption to offer the most cost-effective colony expansion compared to leading competitors.
StemFit® is part of our Stem Cell Synergy Range - use alongside iMatrix laminin-511 E8 fragments as culture substrate and freeze your cells using CELLBANKER cryopreservation media!
Real World Application
Scientists at Keio University Hospital have successfully transplanted iPS cell-derived neural stem/progenitor cells (iPSC-NS/PCs) into a patient, to start the first-in-human clinical study for the treatment of spinal cord injury using iPSC-derived cells. CiRA’s protocol features StemFit cell culture medium with iMatrix-511 MG and STEM CELLBANKER, all available from AMSBIO.
Benefits
- Animal-Origin-Free & feeder-free
- Weekend-free cell culture
- Reduced media consumption
- Reproducible growth rates
- Easy transition from feeder cells
- Superior colony forming efficiency from a single clone
- Compatible with a large number of matrices
Real World Application
Read the interview with Professor Ryuhei Hayashi about this study and the world’s first transplantation of a corneal epithelial sheet-derived from induced pluripotent stem (iPS) cells using StemFit.
The StemFit® Range
Basic03
Human and animal-origin free, defined stem cell culture medium for human pluripotent stem cell culture. Capable of maintaining human ES and iPS cells under feeder-free culture conditions.
Basic04 Complete
Human and animal-origin free, defined stem cell culture medium for human pluripotent stem cell culture. Supplied as a single bottle including bFGF; can be used for maintaining human ES and iPS cells under feeder-free culture conditions.
| Feature | Basic03 | Basic04 Complete |
|---|---|---|
| Suitable for Single-cell Culture | Yes | Yes |
| Free of Animal & Human Material | Yes | Yes |
| cGMP | Available† | Available |
| Number of Bottles | 2 | 1 |
| bFGF | Sold Separately | Included (80ng/ml) |
† SFB-503-GMP: Made in USA under US cGMP
Product Information Table
| Name | Datasheet | Packsize | Order |
|---|---|---|---|
| StemFit® Basic03 - Clinical Grade Stem Cell Culture Media |  | Liquid A: 400ml / Liquid B: 100ml | View |
| StemFit® Basic03 - GMP Grade Stem Cell Culture Media | | Liquid A: 400ml / Liquid B: 100ml | View |
| StemFit® Basic04 Complete - Stem Cell Culture Media including 80ng/ml bFGF, GMP | | 500ml | View |
Weekend-free Cell Culture
The maintenance of stem cells is a complicated and labor-intensive process, requiring a tedious feeding step at the weekends. StemFit® Stem Cell Culture Media frees up your weekends - simply use the recommended weekly workflow shown below to reduce workload and save time!
Fig 1. Suggested weekly workflow using StemFit® Stem Cell Culture Media.
Reduced Media Consumption
Due to the high-quality components and the ideal concentration of nutrients, the required volume of StemFit® is lower than that of conventional media. In addition, the reduced number of feeding steps during the week results in the reduction of media consumption between StemFit® and conventional media by more than 50%.
Fig 2. The volume of StemFit® can be reduced by 25% per well. The reduction in number of media changes per week leads to a further volume reduction of more than 50%.
Reproducible Growth Rate
Our feeder-free media means no more variation due to feeder cells! Cultivating stem cells using StemFit® results in a very reproducible growth rate, meaning you can plan your experiments better. Analyzing the morphology of stem cells cultivated in StemFit® shows that the colony shape and size are very similar to the cells grown on feeder cells.
Total Fold Expansion
Colony Morphology
Fig 3. Human 201B7 iPSCs were cultured on iMatrix Laminin-511 with StemFit® for 4 weeks without weekend feeding. Cell colonies were dissociated into single cells and seeded at the listed densities. (Left) Cell growth rates and density remain consistent regardless of starting seeding densities. (Right) Colony morphology of cells cultured with StemFit®.
Easy Transition from Feeder Cells
The transition from feeder cell conditions to feeder-free could not be easier with StemFit®. Simply switch the medium two days before passaging cells and continue cultivating them in StemFit® feeder-free media. After 2-3 passages the feeder cells will be completely diluted out and you will be left with a pure stem cell culture.
Fig 4. Suggested workflow for transitioning cells to feeder-free media.
Start From A Single Cell
StemFit® Stem Cell Culture Medium allows for a superior colony forming efficiency from a single cell clone, which minimizes the effects of stress and results in reliable cells for downstream applications.
Fig 5. Human iPSCs were adapted to StemFit®, or commercially available medium A or B on Matrigel® for more than 3 passages. Then, cells were serially diluted and seeded with each medium on Matrigel®-coated 96-well plates at 1 cell/well or 10 cells/well. The number of seeded cells was counted after 3 hours, and colonies were counted at day 7.
Greater than 100x Fold Expansion
StemFit® has been tested on many different matrices. There is a greater than 100 fold increase in the expansion rate of cells cultured in StemFit® when compared to Medium A from the market leading competitor. Different matrices were used as a growing matrix and combined with StemFit® or with Medium A.
Fig 6. Human 201B7 iPSCs grown on MEFs (feeder dependent) were transitioned to feeder-free conditions with StemFit® feeder-free media or commercially available medium A on respective ECMs (1000 cells/cm2), and cultured for one week on pre-coated plates.
Consistent Gene Expression
Typical methods of stem cell cultivation can subject cells to unnecessary stress. Unwanted changes in the gene expression profile can be introduced during every passage or growth period. Because of this, the Cell and Gene Therapy Catapult (CGT Catapult) in London investigated the genomic profile of cells cultured in StemFit® compared to four other commercially available media. The most consistent gene expression was obtained using StemFit®. For further details, refer to Superior Performance of StemFit for the Culture of Induced Pluripotent Stem Cells, an independent study performed by CGT Catapult.
Fig 7. Consistent gene expression using StemFit® Media (labeled as AK03N). The expression profile using the TaqMan Scorecard assay (n=3) showed that the most consistent gene expression after 5 passages was obtained using StemFit® media.
Normal Karyotype Analysis after 52 passages
201B7 cells were passaged in StemFit medium and iMatrix-511 substrate for 52 passages. G-band staining showed cells maintained a normal karyotype after 52 passages.
Less Lactate Production
Hypoxic stress can result in the production of lactate which can lead to changes in the genome expression profile or unwanted differentiation of stem cells. The CGT Catapult showed that there is considerably less lactate production when the cells are grown in StemFit®.
Fig 8. StemFit® Media (labeled as AK03N) caused a considerably lower accumulation of lactate compared to 4 other commercially available media.
Clinical Trial Data
Clinical trials using hPSC-derived cells have been initiated recently for various diseases, such as age-related macular degeneration (AMD), Parkinson’s disease (PD), spinal cord injury, and heart failure. Technology to produce quality cells is essential to the success of cell therapy.
Feeder cells and serum-derived components were previously used for the early stages of hPSC culture. However, current methods employ a combination of extracellular matrices (ECMs) and defined xeno-free (XF) and animal-origin-free (AOF) medium under feeder-free conditions to improve safety, consistency, and efficiency. Our AOF, weekend-free StemFit media has been used in several clinical trials alongside our iMatrix-511 as the cell culture substrate.
| ID Number | Year | Cell line | Derived cell type | Indication | Culture medium | Scaffold | Reference |
|---|---|---|---|---|---|---|---|
| UMIN000033564 | 2018 | iPSC (QHJI01s04, episomal) | Dopaminergic progenitors | Parkinson’s disease | AOF medium (StemFit) | Feeder-free (Laminin511E8) | Doi et al. (2020) |
| UMIN000036539 jRCTa050190084 | 2019 | iPSC (YZWJs524, episomal) | Corneal epithelial cell sheet | Limbal stem-cell deficiency | AOF medium (StemFit) | Feeder-free (Laminin511E8) | Hayashi et al. (2017) |
| UMIN000035074 jRCTa031190228 | 2020 | iPSC (YZWJs513, episomal) | Neural stem/progenitor cells | Spinal cord injury | AOF medium (StemFit) | Feeder-free (Laminin511E8) | Itakura et al. (2017) Sugai et al. (2016) |
| jRCTa050190104 | 2020 | iPSC (QHJI01s04, episomal) | Cartilage | Articular cartilage damage | AOF medium (StemFit) | Feeder-free (Laminin511E8) | Yamashita et al. (2020) |
| NCT04945018 | 2021 | iPSC (QHJI14s04, episomal) | Cardiomyocyte spheroids | Heart failure | AOF medium (StemFit) | Feeder-free (Laminin511E8) | Tohyama et al. (2017) |
| NCT04696328 UMIN00032989 | 2021 | iPSC (QHJI14s04, episomal) | Cardiomyocytes sheet | Ischemic cardiomyopathy | AOF medium (StemFit) | Feeder-free (Laminin511E8) | Miyagawa et al. (2022) Kawamura et al.(2016) |
Additional StemFit® products
StemFit Purotein® Recombinant Proteins
Our StemFit Purotein® Recombinant Proteins are a range of easy-to-use animal origin-free growth factors and cytokines. These enable precise differentiation of ESCs and iPSCs into a variety of lineages including liver, pancreas, kidney, heart, blood and immune cells.
StemFit® for Differentiation
StemFit® For Differentiation is a chemically defined and human/animal component-free supplement for differentiation of human ES and iPS cells to multiple lineages. It can be used with a variety of different induction factors or cytokines to support differentiation along ectoderm, mesoderm or endoderm lineages.
StemFit® For Mesenchymal Stem Cells
StemFit® For MSCs is a chemically defineddefned medium optimized for the expansion and maintenance of human bone marrow-derived MSC (BM-MSC), umbilical cord-derived MSC (UC-MSC), and adipose-derived stem cells (ADSC), under serum-free, human platelet lysate free conditions.
StemFit® FAQs
- High fold expansion rate (~100× expansion / weekly passage).
- Reproducible and manageable culture by controlling the numbers of seeded cells.
- Cost-effective culture with lower medium volume and less frequent medium changes.
- Produce an iPSC colony derived from single cells (essential for genome editing).
- Any commercially available bFGF have been confirmed to work. Ajinomoto provides high quality, animal origin-free bFGF (Item code:
SP-FGF2-G-001MG)
- We recommend iMatrix laminin-511 E8 fragments as the superior cell culture substrate for Stemfit media. Any other commercially available ECMs for hPSCs can also work.
- Yes, but it is recommended to make a small clump and seeding at a low cell density.
- We recommend switching to Stemfit 2-3 days prior to passaging.
Please refer to Technical Tips for more details.
- Adjust the bFGF concentration (e.g. 40 - 80 ng/mL) according to your cell line.
- Try a higher seeding density (e.g. > 1.0 x 105 cells per a well of 6-well plate). Distribute the cells evenly upon passage.
- Culture in Y-27632-containing medium for more than 24 hours after passaging.
Make sure that the medium was thawed within 2 weeks and has not been heated to 37°C. - Detached cells with low viability may not grow well.
- For high density-seeding, wells should be pre-coated with ECM. Uncoated wells will lead to poor attachment.
- Precisely count cell number in the cell suspension, calculate volume to seed.
- Cryopreserved cells cannot be seeded by the uncoated method since they are usually seeded in high density
- StemFit medium has been developed for human PSCs, however there are reports in which primate PSCs have been cultured in StemFit medium.
- 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.
- Basic02 and Basic03 have the same composition but the origin of raw material is different. Basic03 is free of human or animal derived ingredients, whereas Basic02 does contain human derived ingredients.
Featured Citations
Neumayer, G. et al (2024). Nature Communications,15(1).
In Vivo Assessment of Laboratory-Grown Kidney Tissue Grafts. Bioengineering.
Chuang, T. et al. (2023).Bioengineering (Basel). 10(11):1261.
Asynchronous mixing of kidney progenitor cells potentiates nephrogenesis in organoids
Gupta, A. K.et al. (2020) Communications Biology 3(1), 1-11.
Human iPSC-Derived Posterior Gut Progenitors Are Expandable and Capable of Forming Gut and Liver Organoids.
Zhang R,R. et al. (2018). Stem Cell Report. 10(3), article number 780–93.
Derivation of induced pluripotent stem cells in Japanese macaque (Macaca fuscata).
Nakai, R. et al. (2018).Scientific Reports. 8, article number 812187.
Optimal Hypoxia Regulates Human iPSC-Derived Liver Bud Differentiation through Intercellular TGFB Signalling.
Ayabe, H. et al. (2018). Stem Cell Reports. Article number 11(2):306–16.
Generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells
Ryuji Morizane & Joseph V Bonventre (2017) Nature Protocols 12, 195–207.
Massive and Reproducible Production of Liver Buds Entirely from Human Pluripotent Stem Cells.
Takebe,T. et al.(2017). Cell Reports. 4, Article number: 2661-70.
Efficient Adhesion Culture of Human Pluripotent Stem Cells Using Laminin Fragments in an Uncoated Manner
Miyazaki, T. et al. (2017) Scientific Reports 7: 41165.
Higher-Order Kidney Organogenesis from Pluripotent Stem Cells. Cell Stem Cell.
Taguchi A, Nishinakamura R. (2017). 21(6):730-746.e6.
Multilineage communication regulates human liver bud development from pluripotency
Camp, J. G. et al. (2017). Nature.22;546(7659):533-538.
Single-cell transcriptome of early embryos and cultured embryonic stem cells of cynomolgus monkeys.
Nakamura, T. et al.(2017).Scientific Data. 4(1).
Generation of a Three-Dimensional Kidney Structure from Pluripotent Stem Cells.
Yoshimura, Y. et al. (2017). Methods in molecular biology. 1;179–93.
Engineering the AAVS1 locus for consistent and scalable transgene expression in human iPSCs and their differentiated derivatives
Oceguera-Yanez F, et al. (2016). 101:43–55.
A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells
Nakagawa, M. et al.(2014). Scientific Reports. 4, Article number: 3594.
Vascularized and functional human liver from an iPSC-derived organ bud transplant.
Takebe, T. et al.(2013). Nature. Article number:499(7459):481–4.
