Product Classification : hES/iPSC Culture Media / ReagentsResearch Theme : ES/iPS cells
Feeder-Free Culture Medium for human ES/iPS cells
- Require culture maintenance only three days per week. (i.e. Monday, Wednesday and Friday)
- Reduces medium usage by 50% compared to medium that require change everyday.
- Equivalent performance to feeder dependent medium in maintaining undifferentiated state.
- Maintain differentiation capability. (differentiated into cadiomyocytes, neurons and others)
- Each batch Serum-free is functionally tested by using human iPS cells. Osmolarity, pH, sterility, and mycoplasma-contamination is tested to ensure batch-to-batch consistency.
Note: bFGF is not included in ReproFF2. Please purchase bFGF separately and add it before you culture human ES cells and iPS cells.
Culture maintenance only on Monday, Wednesday and Friday
With ReproFF2, medium change is not required everyday.
Culture doesn’t have to be cared during the weekend if passaging is done on Friday.
ReproFF2 is an epoch-making feeder-free culture medium for human ES cells and iPS cells. ReproFF2 was developed in order to simplify culture maintenance. While conventional stem cell medium require medium change every day, ReproFF2 requires culture maintenance only three days in a week. For example, you can change medium on Monday and Wednesday, and then passage cells on Friday. Consequently, cost and usage amount of medium will be halved .
*: not required for all cell lines
Human iPS cells (201B7) grown with ReproFF2 on a Laminin-5 coated plate.
Maintenance of an undifferentiated state by using ReproFF2
Cells cultured with ReproFF2 retain the key characteristics of pluripotent cells
Maintanance of undifferentiated human iPS cells with ReproFF2
human iPS cell line 201B7
Feeder-free culture with ReproFF2: Passage 10, day 7
Feeder-dependent culture on MEF: Passage 48, day 3
1. Morphology and ALP activity
Human iPS cell line 201B7 (established in Kyoto University) was kept undifferentiated in feeder-free culture with ReproFF2, as confirmed by morphology (A) and alkaline phosphatase staining (B). These results looked comparable to those of feeder-dependently cultured iPS cells (C and D).
2. Expression of pluripotency markers
Human iPS cells cultured with ReproFF2 retained expression of pluripotency marker genes Oct 3/4, SSEA-4, TRA-1-60 and Nanog, similarly to cells cultured on feeder cells.
3. FACS analysis of pluripotency marker expression in iPS cells
Human iPS cells cultured with ReproFF2 showed profiles comparable to those of feeder-dependently cultured cells when analysed with FACS for pluripotency marker genes SSEA-4, TRA-1-60 and TRA-1-81.
4. Karyotype analysis of human iPS cells after long term culture with ReproFF2
Maintanance of undifferentiated human ES cells with ReproFF2
Protocol using Gentle Cell Dissociation Reagent（STEM CELL TECHNOLOGIES, 07174）and Vitronectin-N（LIFE TECHNOLOGIES, REFA14700）
Protocol using Dissociation Solution (RCHETP002) and Laminin-5 (RCHEOT004) / Matrigel (Corning)
Safety Data Sheet
- Morizane R, et al. “Nephron organoids derived from human pluripotent stem cells model kidney development and injury.” Nat Biotechnol. 2015 Oct 12. doi: 10.1038/nbt.3392.
- Naoki Nishishita, et al. “Generation and Maintenance of iPSCs From CD34+Cord Blood Cells on Artificial Cell Attachment Substrate.” Biochemistry, Genetics and Molecular Biology, Chapter 5. http://dx.doi.org/10.5772/58591
- Kanemura, Hoshimi, et al. “Tumorigenicity Studies of Induced Pluripotent Stem Cell (iPSC)-Derived Retinal Pigment Epithelium (RPE) for the Treatment of Age-Related Macular Degeneration.” PloS one 9.1 (2014): e85336.
- Hasegawa, Tsuyoshi, et al. “Cancer‐associated fibroblasts might sustain the stemness of scirrhous gastric cancer cells via transforming growth factor‐β signaling.” International Journal of Cancer (2013).
- Kanemura, Hoshimi, et al. “Pigment epithelium-derived factor secreted from retinal pigment epithelium facilitates apoptotic cell death of iPSC.” Scientific reports 3 (2013).
- Lam, Albert Q., et al. “Rapid and Efficient Differentiation of Human Pluripotent Stem Cells into Intermediate Mesoderm That Forms Tubules Expressing Kidney Proximal Tubular Markers.” Journal of the American Society of Nephrology: JASN (2013).
- Murakami, Masashi, et al. “The use of granulocyte-colony stimulating factor induced mobilization for isolation of dental pulp stem cells with high regenerative potential.” Biomaterials 34.36 (2013): 9036-9047.
- Nishishita, Naoki, et al. “Generation of virus-free induced pluripotent stem cell clones on a synthetic matrix via a single cell subcloning in the naive state.” PloS one 7.6 (2012): e38389.
- Sugiyama, Norikazu, et al. “Label-free characterization of living human induced pluripotent stem cells by subcellular topographic imaging technique using full-field quantitative phase microscopy coupled with interference reflection microscopy.” Biomedical optics express 3.9 (2012): 2175-2183.
Feeder-Free Culture Reagents
|RCHEMD006A||ReproFF2 (500mL) x3||Ask|
|RCHEMD006B||ReproFF2 (500mL) x3 & bFGF(25μL) pack||Ask|
Feeder-Dependent Culture Reagents
|RCHEMD001||Primate ES cell medium (500mL)||Ask|
|RCHEMD001A||Primate ES cell medium (500mL) x5||Ask|
|RCHEMD001B||Primate ES cell medium (500mL) x5 & bFGF(25μL) pack||Ask|
|RCHEMD005A||ReproStem (500mL) x5||Ask|
|RCHEMD005B||ReproStem (500mL) x5 & bFGF(25μL) pack||Ask|
|RCHECK002||Primate ES cell culture kit||Ask|
Feeder-Free Culture Reagents
|RCHEMD004A||ReproFF (500mL) x5||Ask|
|RCHEMD004B||ReproFF (500mL) x5 & bFGF(25μL) pack||Ask|
Othe Culture Reagents
|RCHETP002||Dissociation solution for human ES/iPS cells (30mL)||Ask|
|RCHEFM001||Freezing medium for human ES/iPS cells (25mL)||Ask|