Benefits

StemFit® Basic03 has been used together with iMatrix-511 recombinant laminin E8 in a world-first clinical study of iPSC-derived corneal epithelial cells by Professor Ryuhei Hayashi and co-workers from the Graduate School of Medicine, Osaka University, Japan. See more details and an interview here.

Stem Cell Synergy: Grow and Store

AMSBIO's iMatrix laminin can be used together with our StemFit cell culture media and CELLBANKER series cryopreservation solution, to create the perfect Stem Cell Synergy for your ES/iPSC cell culture!

The StemFit® Range

Basic03

Human and animal-origin free, defined 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 medium for human pluripotent stem cell culture. Supplied as single bottle including bFGF; can be used for maintaining human ES and iPS cells under feeder-free culture conditions.

FeatureBasic03Basic04 Complete
Suitable for Single-cell CultureYesYes
Free of Animal & Human MaterialYesYes
cGMPAvailable†Available
Number of Bottles21
bFGFSold SeparatelyIncluded (80ng/ml)

SFB-503-GMP: Made in USA under US cGMP

Product Information Table

NameDatasheetPacksize Order
StemFit Basic03 (animal-origin free, defined medium for human pluripotent stem cell culture)Liquid A:400 ml/Liquid B:100 ml View
StemFit ® Basic03 GMP grade (animal-origin free, defined medium for human pluripotent stem cell culture, GMP grade)Liquid A:400 ml/Liquid B:100 ml View
StemFit® Basic04 Complete (animal-origin free, defined medium for human pluripotent stem cell culture, including 80ng/ml bFGF)500 ml View
StemFit® Basic04 Complete GMP Grade (animal-origin free, defined medium for human pluripotent stem cell culture, including 80ng/ml bFGF, GMP Grade)-500 ml 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® frees up your weekends - simply use the recommended weekly workflow shown below to reduce workload and save time!

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. Additionally, reduced feeding steps during the week, creates a total volume difference between StemFit® and conventional media of more than 50%.

Conventional Protocol vs StemFit protocol

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

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. Not only that, 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
Colony Morphology

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.

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®. After 2-3 passages the feeder cells will be completely diluted out and you will be left with a pure stem cell culture.

Start From A Single Cell

StemFit® 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.

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.

Consistent Gene Expression

Typical methods of stem cell cultivation can subject cells to a lot of 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 4 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.

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®.

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 NumberYearCell lineDerived cell typeIndicationCulture mediumScaffoldReference
UMIN0000335642018iPSC (QHJI01s04, episomal)Dopaminergic progenitorsParkinson’s diseaseAOF medium (StemFit)Feeder-free (Laminin511E8)Doi et al. (2020)
UMIN000036539 jRCTa050190084 2019iPSC (YZWJs524, episomal)Corneal epithelial cell sheetLimbal stem-cell deficiencyAOF medium (StemFit)Feeder-free (Laminin511E8)Hayashi et al. (2017)
UMIN000035074 jRCTa0311902282020iPSC (YZWJs513, episomal)Neural stem/progenitor cellsSpinal cord injuryAOF medium (StemFit)Feeder-free (Laminin511E8)Itakura et al. (2017)
Sugai et al. (2016)
jRCTa0501901042020iPSC (QHJI01s04, episomal)CartilageArticular cartilage damageAOF medium (StemFit)Feeder-free (Laminin511E8)Yamashita et al. (2020)
NCT049450182021iPSC (QHJI14s04, episomal)Cardiomyocyte spheroidsHeart failureAOF medium (StemFit)Feeder-free (Laminin511E8)Tohyama et al. (2017)
NCT04696328 UMIN000329892021iPSC (QHJI14s04, episomal)Cardiomyocytes sheetIschemic cardiomyopathyAOF medium (StemFit)Feeder-free (Laminin511E8)Miyagawa et al. (2022)
Kawamura et al.(2016)

Additional StemFit® products

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 defned 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 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® FAQs

What are the benefits of single cell culture? / Why is single cell culture recommended?
  • 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).
Which bFGF can be used?
  • Any commercially available bFGF have been confirmed to work. Ajinomoto provides high quality, animalorigin free bFGF (Item code: SP-FGF2-G-001MG)
Which Extracellular Matrix (ECM) is recommended?
  • 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.
Can I use StemFit for clump culture?
  • Yes, but it is recommended to make a small clump and seeding at a low cell density.
How can I switch my medium to Stemfit?
  • We recommend to switch to Stemfit 2-3 days prior to passaging.
    Please refer to Technical Tips for more details.
Cells do not grow well?
  • 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 hoursafter 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.
  • Uncoated method cannot be applied high density-seeding. (Cells poorly attach to the wells).
Cells do not attach to the wells.
  • 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
Can I use StemFit 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.
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.
What’s the difference between Basic03 and discontinued Basic02?
  • Basic02 and Basic03 has the same composition but the origin of raw material is different. Basic03 is free of human or animal derived ingredient, whereas  Basic02 does contain human derived ingredient.

Featured Citations

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.

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.

Generation of a Three-Dimensional Kidney Structure from Pluripotent Stem Cells.
Yoshimura, Y. et al. (2017). Methods in molecular biology. 1;179–93.

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.