StemFit® Feeder-Free Stem Cell Culture Media
Allowing weekend-free culture of stem cells
StemFit® is a range of xeno-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 and growth medium.
Available in research and clinical grade formulations, StemFit® 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.
- Xeno-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
The StemFit® Range
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.
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.
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)
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 Differentiation can be combined with StemFit® Basic media for human PSCs, to support clinical research applications of hPSC-derived cells/tissues by providing defined and xeno-free culture systems for both expansion and differentiation. StemFit® For Differentiation is provided as 5X concentrate and is intended to be used with basal medium (DMEM, DMEM/F12, RPMI1640, etc.) and appropriate induction factors or cytokines.
StemFit® For Mesenchymal Stem Cells (MSC)
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.
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%.
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
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.
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®.
- High fold expansion rate (~100× expansion / weekly passage).
- Reproducible and manageable culture by controlling the numbers of seeded
- Cost-effective culture with lower medium volume and less frequent medium changes.
- Produce an iPSC colony derived from single cells (essential for genome editing).
- Yes, but it is recommended to make a small clump and seeding at a low cell density.
- Any commercially available bFGF have been confirmed to work. Ajinomoto provides high quality, animalorigin free bFGF (Item code: SP-FGF2-G-001MG)
- 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).
- 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
Asynchronous mixing of kidney progenitor cells potentiates nephrogenesis in organoids
Gupta, A. K., Sarkar, P., Wertheim, J. A., Pan, X., Carroll, T. J., & Oxburgh, L. (2020) Communications Biology 3(1), 1-11.
Generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells
Ryuji Morizane & Joseph V Bonventre (2017) Nature Protocols 12, 195–207.
Efficient Adhesion Culture of Human Pluripotent Stem Cells Using Laminin Fragments in an Uncoated Manner
Miyazaki, T. et al. (2017) Scientific Reports 7: 41165.
Multilineage communication regulates human liver bud development from pluripotency
Camp, J. G. et al. (2017). Nature 2017 Jun 22;546(7659):533-538.
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.