Novoprotein Organoid Course: Noggin as A Key Factor in Organoid Culture

Role of Noggin in Regulating BMP and Wnt Signaling During Development

Noggin has a wide range of expression and function in vertebrates, and plays an important role in early embryonic development, limb formation and nervous system development. Its main mechanism of action is that it can specifically bind to and inhibit the activity of bone morphogenetic proteins (BMPs), thus regulating the normal development of the organism.

Noggin blockade of BMP signaling leads to facial developmental defects^[1]

BMPs belong to the transforming growth factor-β (TGF-β) superfamily, which is a pleiotropic cytokine widely found in various tissues and organs that regulates cell growth and differentiation, and has an indispensable role in embryonic development, cell differentiation, and regenerative repair. As an inhibitor of bone morphogenetic proteins (BMPs), Noggin regulates cell differentiation, proliferation and apoptosis during development by binding to cytokines such as BMP-2/4/5/6/7 and growth and differentiation factor (GDF)-5/6 with different affinities. It has been shown that Noggin gene has a negative feedback inhibition of the BMP signaling pathway and a positive feedback activation of the Wnt signaling pathway, and that Noggin's modulation of Wnt signaling is likely to be through the BMP signaling pathway to produce a similar effect in regulating cell growth^[2].

In addition, Noggin has been found to promote bone metastasis in certain cancers and has been associated with the development of primary bone malignancies.

Noggin and Organoid Culture

Growth factors, as active components necessary for cell culture, proliferation and differentiation, are essential for the successful establishment of organoids and are one of the important components to ensure sustainable cell culture in organoids. Cytokine combinations commonly used in intestinal models are WNER (Wnt-3a, Noggin, EGF and R-Spondin), in which Noggin has an important position.

First, Noggin and fibroblast growth factor (bFGF) synergistically inhibit the BMP pathway, which helps to maintain human embryonic stem cells (hESCs) in proliferative cultures and stage-stop their spontaneous differentiation^[3]. Secondly, Noggin also binds to BMP4, BMP7, etc., thereby positively regulating Wnt signaling to activate stem cells, and it has been confirmed that long-term culture of liver organoids requires the maintenance of high levels of Wnt signaling as well as sustained inhibition of BMP signaling^[4].

As one of the most basic organoid culture factors, the addition of Noggin cytokines is required in the culture of a variety of normal and tumor organoids, such as intestinal organoids, gastric organoids, lung organoids, prostate organoids, mammary organoids, as well as colorectal carcinoma organoids, pituitary tumor organoids, breast carcinoma organoids, ovarian carcinoma organoids, and so on.

Novoprotein provides  recombinant human or mouse Noggin proteins with low endotoxin, high activity, and high lot-to-lot consistency, and has been validated in multi organoid models, such as human breast cancer organoids, ipsc-derived human small intestinal organoids, and mouse intestinal organoids, colonic organoids, gastric organoids and pancreatic organoids etc. Making your organoid culture more controllable and stable!

Product data

01 High activity

Measured by its ability to inhibit BMP-4-induced alkaline phosphatase production by ATDC5 mouse chondrogenic cells.The ED50 for this effect is 10-100 ng/ml in the presence of 300 ng/mL of Recombinant Human BMP4(Cat#CR93) (QC Verified）

The activity of Recombinant Human Noggin Protein(Cat#CB89) was compared to another commercially available product.

02 High lot-to-lot consistency

Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of Noggin（Cat#CB89）

03 Validated by organoid culture

• ipsc-derived human small intestinal organoids

Human intestinal organoids (ipsc source) were cultured with Activin A(Cat#C687) , BMP-4(Cat#CR93) , EGF (Cat#C029) , Wnt3a (Cat#C22R), Noggin (Cat#C028), R-spondin 1 (Cat#CX83) and FGF-4 (Cat#CR08) . The organoids showed good morphology.

• Breast cancer organoids

Human breast cancer organoids were cultured with Wnt3a(Cat#C22R), Noggin (Cat#CB89)R-spondin 3 (Cat#C18C), KGF (Cat#CH73), FGF-10 (Cat#CR11), EGF (Cat#C029) and NRG1-beta 1 (Cat#C753). DAPI (blue), Ki67(red) and ER(green).

• Mouse intestinal organoids

Mouse intestinal organoids were cultured with EGF (Cat#C029), Noggin (Cat#C028) and R-spondin 1 (Cat#CX83). After five days of culture, the organoids showed good morphology and germination rate.

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References:

[1]Lee, SH., Fu, K., Hui, J. et al. Noggin and retinoic acid transform the identity of avian facial prominences. Nature 414,909–912(2001).

[2]Walsh DW, Godson C, Brazil DP, Martin F. Extracellular BMP-antagonist regulation in development and disease: tied up in knots. Trends Cell Biol. 2010 May;20(5):244-56.

[3]MA Yunan, YOU Ying, SHEN Huanhuan, SUN Zhaizeng, ZENG Lin, FA Yunzhi.Effect of Noggin gene silencing on the expression of BMP and Wnt signaling pathway[J]. Chinese Journal of Laboratory Animal Science, vol,2016,24(5):475-480

[4]Schille, C., Bayerlova, M., Bleckmann, A., and Schambony, A. (2016). Ror2 signaling is required for local upregulation of GDF6 and activation of BMP signaling at the neural plate border. Development 143, 3182–3194.