MBL Cell Culture: Organoid/Stem cell
Factors for Organoid and Stem Cell Culture
MBL International supports advances in cell culture by offering Afamin/Wnt3a Condition Medium and recombinant Afamin/Wnt3a.
Wnt signaling is known to be involved in early development, maintenance and regeneration of stem cells, and in cancer formation. Wnt signaling has also been found to play an important role in the growth and maintenance of these processes. In particular, Wnt3a has been revealed to be an essential niche component for maintaining the proliferation of Lgr5-positive stem cells in intestinal epithelial cells and is used for the production of various digestive organoids such as the small intestine, large intestine, stomach, pancreas and liver. Although Wnt3a has been conventionally used for the culture of gut organoids, it is a fat-soluble protein, so it forms aggregates in serum-free medium and can not exert its activity sufficiently. In 2016, Mihara et al. found that high Wnt3a activity can be maintained by forming a complex with Wnt3a by Afamin, which is one of the components of serum. In addition, by using Afamin and Wnt3a complex for organoid culture, long-term culture of organoid becomes possible. This new medium will result in optimal success for your organoid experiments.
Brochures:
| Product Type |
Product Name (click for order info) |
Size |
CBU SKU |
| Conditioned Medium |
Afamin/Wnt3a CM (Conditioned Medium) |
10 ml |
MBL-J2-001 |
| |
DESCRIPTION: Serum free conditioned medium including Wnt3a-Afamin complex for intestinal organoid cell culture.
Conditioned medium containing Wnt3a protein and Afamin protein. Mouse Wnt3a and the human Afamin were co-expressed in Expi293F™ Cells*. For the co-expression of Wnt3a and Afamin, expression plasmids for N-terminal PA tagged mouse wnt3a and N-terminal TARGET tagged human afamin were co-transfected using the Expi293 Expression System*. Conditioned medium was prepared using serum-free media (Advanced DMEM/F-12* containing streptomycin, HEPES and GlutaMAXTM Supplement*). Then, it was sterilized by filtration of 0.2 µm-filter. FBS was used for pre-culture of Afamin/Wnt3a expression Expi293F™ Cells. *: Thermo Fisher Scientific’s products.
DIRECTION for USE: As a canonical Wnt signal agonist. For intestinal organoids culture, add to the culture media at a final concentration of 10%. Since the optimal condition may vary depending on samples, each user should determine the optimal volume of addition in advance. Formerly Product Code: J-ORMW301R
Note Regarding Intellectual Property Rights: When culturing organoids, or stem cells, or other tissues, if you are to use this product in combination with other factor or factors (hereunder factors), a third party may have a patent on the use or other application of the factors concerned. Regarding to this product, we do not offer any non-infringement warranty when used or otherwise applied in combination with other factors. Therefore, if you intend to use this product in combination with other factors, please check with your organization’s division responsible for intellectual property rights or your research agency before using this product.
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| Conditioned Medium |
Afamin/Wnt3a CM (High Concentration) |
5 ml |
MBL-J2-005-05 |
| |
DESCRIPTION: Conditioned medium (CM) containing Wnt3a protein and Afamin protein.
The mouse Wnt3a and the human Afamin were co-expressed in Expi293F™ Cells*. For the co-expression of Wnt3a and Afamin, expression plasmids for N-terminal PA tagged mouse wnt3a and N-terminal TARGET tagged human afamin were co-transfected using the Expi293 Expression System*. Conditioned medium was prepared using serum-free media (Advanced DMEM/F-12* containing streptomycin, HEPES and GlutaMAXTM Supplement*). Then, it was sterilized by filtration of 0.2 µm-filter. FBS was used for pre-culture of Afamin/Wnt3a expression Expi293F™ Cells. *: Thermo Fisher Scientific’s products.
DIRECTION for USE: As a canonical Wnt signal agonist. For intestinal organoids culture, add to the culture media at a final concentration of 2.5%. Since the optimal condition may vary depending on samples, each user should determine the optimal volume of addition in advance.
Note Regarding Intellectual Property Rights: When culturing organoids, or stem cells, or other tissues, if you are to use this product in combination with other factor or factors (hereunder factors), a third party may have a patent on the use or other application of the factors concerned. Regarding to this product, we do not offer any non-infringement warranty when used or otherwise applied in combination with other factors. Therefore, if you intend to use this product in combination with other factors, please check with your organization’s division responsible for intellectual property rights or your research agency before using this product.
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| Recombinant Protein |
Recombinant Afamin/Wnt3a |
60 ug |
MBL-J2-002 |
| |
DESCRIPTION: Recombinant Wnt3a protein and Afamin protein. The N-terminal PA tagged mouse Wnt3a and the N-terminal Myc tagged human Afamin were co-expressed in CHO-K1 cells.
USE NOTES: We recommend the addition of albumin to dilution buffer to improve freeze-thaw stability, not diluted without albumin such as PBS only, if diluted product is stored in freeze. Dilutions prepared by the following protocol have been shown to maintain high Wnt3a activity after a freeze-thaw cycle.
1. Prepare PBS containing 0.04% HSA (human serum albumin) or BSA (bovine serum albumin).
2. Dilute Recombinant Afamin/Wnt3a with 0.04% HSA or BSA/PBS to prepare 2-, 10-, and 50-fold dilutions.
Use the prepared dilutions as soon as possible and avoid repeated freeze-thaw cycles.
Note Regarding Intellectual Property Rights: When culturing organoids, or stem cells, or other tissues, if you are to use this product (J2-001 Afamin/Wnt3a CM) in combination with another factor or factors (hereinafter “factors”), a third party may have patent rights concerning the use or other application of the factors concerned. MBL does not offer any non-infringement warranty for J2-001 Afamin/Wnt3a CM when used or otherwise applied in combination with other factors. Therefore, if you intend to use this product in combination with other factors, please check with your organization’s division or advisors responsible for intellectual property rights or with your research agency before using this product with other factors.
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| Recombinant Protein |
FGF-Max (Recombinant human FGF-1/FGF-2 chimera) |
50 ug |
MBL-J2-003 |
| |
USE NOTES:
RECONSTITUTION: Reconstitute with 250 µL of ice-cold distilled water on ice.
EXPIRATION: The expiration date is indicated on the tube label. After reconstitution, it can be stable for 1 week at 4°C. Or, prepare appropriate aliquots and freeze them at -80°C using low retention tube. Avoid repeated freezing and thawing.mbinant chimeric protein of FGF-1 (FGF-acidic) and FGF-2 (FGF-basic) expressed in E. coli.
Note Regarding Intellectual Property Rights: If you are to use this product in combination with other factor or factors (hereunder factors) when culturing organoids, stem cells, or other tissues, a third party may have a patent on the use or other application of the factors concerned.
Regarding this product, we do not offer any non-infringement warranty when used or otherwise applied in combination with other factors. Therefore, if you intend to use this product in combination with other factors, please check with your organization’s division responsible for intellectual property rights or your research agency before using this product.
Find more information about FGF-Max here.
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| Recombinant Protein |
Recombinant Human R-spondin 1 |
50 ug |
MBL-J2-004 |
| |
DESCRIPTION: R-spondin 1 (roof plate-specific spondin 1), also known as Christine 3, is one of the R-spondin family members that regulates to the activation of canonical WNT/β-catenin signaling. In the absence of R-spondin, Frizzled (Fz) is endocytosed by being ubiquitinated by the binding of the ubiquitin ligase RNF43/ZNRF3, Fz on the cell membrane decreases, and Wnt signals are not activated enough. Conversely, in the presence of R-spondin, RNF43/ZNRF3 forms trimers with LGR and R-spondin and is endocytosed, making it unable to ubiquitinate Fz. As a result, Fz on the cell membrane increases, and Wnt signals are strongly activated.
This product is a bioactive domain consisting of 21 to 146 amino acids which contain two adjacent cysteine-rich furin-like domains (Fu1 and Fu2). Fu1 binds to the extracellular domain of ZNRF3*1 and RNF43*2, and Fu2 binds to the extracellular domain of LGR4/5/6.
Recombinant protein of human R-spondin 1 (21-146 aa) is expressed in E. coli.
*1 zinc and ring finger 3, *2 ring finger protein 43, *3 leucine-rich repeat-containing G-protein coupled receptors 4/5/6
Organoid culture is typically utilized niche factors that act on Wnt, EGF, BMP and Notch signals that control stem cell differentiation. Among these signals, R-spondin 1 contributes to the activation of Wnt signals.
Note Regarding Intellectual Property Rights: When culturing organoids, stem cells, or other tissues, if you are to use this product in combination with other factor or factors (hereunder factors), a third party may have a patent on the use or other application of the factors concerned. Regarding to this product, we do not offer any non-infringement warranty when used or otherwise applied in combination with other factors. Therefore, if you intend to use this product in combination with other factors, please check with your organization’s division responsible for intellectual property rights or your research agency before using this product.
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