Optimizing Organoid Culture Conditions
Identifying and optimizing the conditions for culturing organoids can be technically challenging. Organoid cultures typically contain multiple components and can be highly variable for different types of organoids. As a result, it can take time, and a lot of trial and error to find the appropriate conditions to support robust organoid growth. Some of the most important components of organoid media are growth factors, which need to be highly active and display minimal variability from lot-to-lot to ensure that they promote optimal organoid growth and help establish reproducible culture conditions.
R&D Systems is a trusted source for recombinant proteins for organoid research. Our stringent production and purification standards ensure that our proteins will provide superior performance and batch-to-batch consistency. This allows researchers to be confident that their growth factors will not only support robust organoid growth, but that their culture conditions will be reproducible from one experiment to the next.
Select Recombinant Proteins for Organoid Research
Advantages of R&D Systems Proteins for Culturing Organoids
High level of bioactivity
Bioactivity testing is performed to ensure that all of our proteins display high levels of activity. Our Recombinant Human Noggin, R-Spondin 1, and Wnt-3a proteins consistently display higher levels of activity than leading competitors’ proteins.
Lot-to-lot consistency
Our proteins undergo rigorous testing to ensure that each new lot displays the same level of bioactivity and purity as previous lots, so researchers can trust that their experiments will be reproducible.
Ability to produce even the most complex proteins
Some proteins commonly used for culturing organoids, such as Recombinant Human Noggin, R-Spondins, and Wnt proteins can be difficult to express and purify. With over 30 years of experience in protein production and purification, we have the expertise needed to purify even the most difficult proteins.
Low endotoxin levels
Each new lot is tested to make sure that it meets our expectations for endotoxin specifications, which is an industry-leading <0.1 EU/µg.
Highly published
A high number of publications cite the use of R&D Systems recombinant proteins for organoid culture, including Recombinant Human R-Spondin 1, R-Spondin 3, Noggin, and Wnt-3a proteins.
Stability of source
We have the capacity and manufacturing experience to ensure that we can provide you with the products you need, when you need them.
Single point of contact for other organoid culture reagents
In addition to our recombinant proteins, we also offer a wide selection of other reagents for culturing organoids, including:
- Organoid Qualified Cultrex® Reduced Growth Factor (RGF) Basement Membrane Extract (BME), Type 2 for robust organoid cultures and Type R1 for difficult organoid cultures
- N-2 MAX and N21-MAX Media Supplements
- Small molecules commonly used as 3-D growth matrix components
- Cryopreservation media for optimized cryopreservation of cell lines or stem cells
- Cultrex Organoid Harvesting Solution
- R&D Systems antibodies and RNAScope® in situ hybridization assays for identifying key lineage-specific markers
Culture and Characterization of Adult Stem Cell-derived Human Liver and Descending Colon Organoids
Examples of Bioactivity and Lot-to-Lot Consistency Testing of Proteins for Culturing Organoids
Recombinant Human Noggin:
R&D Systems Recombinant Human Noggin is 30-fold More Active than Recombinant Human Noggin from a Leading Competitor and Displays Minimal Lot-to-Lot Variability. (A) Using the ATDC5 mouse chondrogenic cell line, the ability of R&D Systems Recombinant Human Noggin (Catalog # 6057-NG; orange line) or a top competitor’s recombinant human noggin (blue line) to inhibit alkaline phosphatase production stimulated by Recombinant Human BMP-4 (R&D Systems, Catalog # 314-BP; 50 ng/mL) was assessed. The ED50 for R&D Systems Recombinant Human Noggin using this assay was approximately 30-fold higher than Noggin obtained from a leading competitor. (B) Using the same assay described in part (A), the lot-to-lot consistency of R&D Systems Recombinant Human Noggin (Catalog # 6057-NG) was assessed by side-by-side testing of the bioactivity of three independent lots of the protein (purple, light blue, orange trace on the graph). The blue trace on the graph is showing the ability of Recombinant Human BMP-4 (R&D Systems, Catalog # 314-BP; 50 ng/mL) to induce alkaline phosphatase production in the ATDC5 mouse chondrogenic cell line.
Recombinant Human R-Spondin 1:
R&D Systems Recombinant Human R-Spondin 1 is 7-fold More Active than Recombinant Human R-Spondin 1 from a Leading Competitor and Displays Minimal Lot-to-Lot Variability. (A) Using a TOPflash beta-Catenin/TCF reporter assay in the HEK293T human kidney cell line, the activity of R&D Systems Recombinant Human R-Spondin 1 (Catalog # 4645-RS; blue line) or a top competitor’s recombinant human R-Spondin 1 (purple line) was assessed, in the presence of Recombinant Mouse Wnt-3a (R&D Systems, Catalog # 1324-WN; 5 ng/mL). The ED50 for R&D Systems Recombinant Human R-Spondin 1 using this assay was approximately 7-fold higher than the leading competitor’s R-Spondin 1. (B) Using the same assay described in part (A), the lot-to-lot consistency of R&D Systems Recombinant Human R-Spondin 1 (Catalog # 4645-RS) was assessed by side-by-side testing of the bioactivity of three independent lots of the protein.
Recombinant Human Wnt-3a:
R&D Systems Recombinant Human Wnt-3a is 1.7-fold More Active than Recombinant Human Wnt-3a from a Leading Competitor and Displays Minimal Lot-to-Lot Variability. (A) Using the MC3T3-E1 mouse preostoblast cell line, induction of alkaline phosphatase production by either Recombinant Human Wnt-3a from R&D Systems (Catalog # 5036-WN; orange line) or from a top competitor (blue line) was assessed. The ED50 for R&D Systems Recombinant Human Wnt-3a using this assay was approximately 1.7-fold higher than Wnt-3a obtained from a leading competitor. (B) Using the same assay described in part (A), the lot-to-lot consistency of R&D Systems Recombinant Human Wnt-3a (Catalog # 5036-WN) was assessed by side-by-side testing of the bioactivity of three independent lots of the protein.
Protein Characterization Using SEC-MALS Analysis
A. Recombinant Human R‑Spondin 3 Protein SEC-MALS. Recombinant Human R-Spondin 3 (Catalog # 3500-RS) has a molecular weight (MW) of 36.1 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
| SEC-MALS Data | Result |
| Retention Time | 12.6-13.1 min |
| MW-Predicted (Monomer) | 28.3 kDa |
| MW-MALS | 36.1 kDa |
| Polydispersity | 1.004 |
| System Suitability: BSA Monomer 66.4 ± 3.32 kDa | Pass |
B. Recombinant Human Noggin Protein SEC-MALS. Recombinant Human Noggin (Catalog # 6057-NG) has a molecular weight (MW) of 58.9 kDa as analyzed by SEC-MALS, suggesting that this protein is a homodimer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
| SEC-MALS Data | Result |
| Retention Time | 12.6-13.2 min |
| MW-Predicted (Monomer) | 23.0 kDa |
| MW-MALS | 58.9 kDa |
| Polydispersity | 1.002 |
| System Suitability: BSA Monomer 66.4 ± 3.32 kDa | Pass |
C. Recombinant Human FGF basic/FGF2/bFGF (145 aa), Tissue Culture Grade Protein SEC-MALS. Recombinant Human FGF basic/FGF2/bFGF, 145 aa TC Grade (Catalog # 4114-TC) has a molecular weight (MW) of 17.2 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
| SEC-MALS Data | Result |
| Retention Time | 16.0-16.6 min |
| MW-Predicted (Monomer) | 16.0 kDa |
| MW-MALS | 17.2 kDa |
| Polydispersity | 1.001 |
| System Suitability: BSA Monomer 66.4 ± 3.32 kDa | Pass |
Related Information
Organoid and 3-D Cell Culture Protocols
R&D Systems scientists are working hard to develop and optimize the culture conditions for growing, expanding, and passaging different types of organoids. Use the protocols that we have developed to simplify your research.
Optimizing Organoid Culture Conditions: The Importance of Growth Factor Bioactivity and Reagent Consistency
View this application note to learn how R&D Systems proteins can help organoid researchers be confident their culture conditions are consistent and reproducible from one experiment to the next.
Cultrex® Basement Membrane Extracts and Extracellular Matrices
Explore our selection of Cultrex Basement Membrane Extract (BME) for robust and difficult to culture organoids. Learn how these products can improve your organoid cultures.
Custom Proteins in a Plate
Our Custom Proteins in a Plate service allows researchers to design your own ready-to-use, custom 96-well plate with the proteins of your choice. Select from a library of over 5,000 bioactive proteins, and our scientists will aliquot and map them for you in a 96-well plate to your specifications.
Evolution of Model Systems for Toxicology, Drug Screening, and Disease Modeling eBook
Read this eBook to gain a greater understanding of the history, current state, and future use of cell-based 2-D and 3-D model systems for toxicology, drug screening, and disease modeling.
Organoids and Beyond – 3-D Tissue in a Dish webinar
Learn how researchers are using organoids to study the basic biology of organ systems and explore regenerative and personalized medicine. Topics discussed in this webinar include protocols for building brain and intestinal organoids and the challenges of using organoids to recapitulate in vivo complexity.