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Recombinant human/rat/bovine/porcine FGF-10 protein (Qk003)

Human/rat/porcine/bovine fibroblast growth factor 10 (FGF10) protein promotes lung organoid formation and induces branching morphology.  FGF10 protein is used widely in organoid culture, embryonic stem cell (ESC) and induced-pluripotent stem cell (iPSC) differentiation, and for the study of epithelial to mesenchymal transition (EMT) and tumor metastasis.

Qkine Qk003 is a high purity and bioactivity 17 kDa bioactive domain of human FGF10, animal origin-free (AOF) and carrier-protein free (CF).

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1000µg will be despatched as 2 x 500µg

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Summary

  • High purity human FGF10 protein (residues 64-208, Uniprot: O15520)

  • >98%, by SDS-PAGE quantitative densitometry

  • 17 kDa

  • Expressed in E. coli

  • Animal origin-free (AOF) and carrier protein-free

  • Manufactured in our Cambridge, UK laboratories

  • Lyophilized from HEPES/NaCl/mannitol

  • Resuspend in water at >100 µg/ml, prepare single use aliquots, add carrier protein if desired and store frozen at -20oC or -80oC

Featured applications

  • Differentiation of embryonic stem cells into gut-like structures, cardiomyocytes and hepatocytes

  • Epithelial to mesenchymal transition

Fibroblast Growth Factor 10, FGFA, KGF2, Keratinocyte growth factor 2

human, rat, bovine, porcine

species similarity:
mouse – 94%

Bioactivity

Human FGF-10 Qk003 bioactivity EMT immunofluorescence lot #010

FGF-10 activity is determined using the firefly luciferase reporter assay in stably transfected HEK293T cells. Cells are treated in triplicate with a serial dilution of FGF-10. Firefly luciferase activity is measured and normalized. EC50 = 21.1 pM (0.36 ng/mL).Data from Qk003 lot #104403.

Purity

Human FGF-10 Qk003 protein purity SDS-PAGE lot #010

FGF10 migrates as a single band at 17 kDa in non-reducing (NR) conditions and upon reduction (R). No contaminating protein bands are visible. Purified recombinant human FGF10 protein (7 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced conditions (NR) and stained with Coomassie Brilliant Blue R250. Data from Qk003 lot #010.

Further quality assays

  • Mass spectrometry: single species with expected mass

  • Endotoxin: <0.005 EU/μg protein (below level of detection)

  • Recovery from stock vial:  >95%

We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine.  All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Protein background

The mature form of human FGF10 protein is an approximately 20 kDa protein highly similar to FGF7 and with a serine-rich region near its N-terminus [1]. It is secreted by mesenchymal cells and is bound and activated by extracellular FGF-BP [2]. Human FGF10 protein is expressed in the mesenchyme and functions through interacting with the epithelial FGF Receptor 2b (FGFR 2b) [3]. It has also been shown to interact weakly with FGFR 1b [4].

Human fibroblast growth factor 10 is first active in the limb bud mesoderm where it creates and maintains FGF signaling with epithelial FGF8, then drives a positive feedback loop accumulating mesenchyme in the growing bud, and finally induces the apical ectodermal ridge which ultimately gives rise to feet and hands [5]. Lung development is based on the same epithelial-mesenchymal FGF mediations involving FGF10 from the foregut mesenchyme signaling to FGFR2 in the foregut epithelium [6]. Furthermore, FGF10 protein is involved in the development of white adipose tissue, heart, liver, brain, kidney, thymus, inner ear, tongue, trachea, eye, prostate, salivary gland and mammary gland. It has been shown to induce migration and invasion of pancreatic cancer cells and to be associated with breast cancer risk, and patients with FGF10 haploinsufficiency present symptoms of chronic obstructive pulmonary disease [3].

FGF10 is involved in a number of different embryo and adult cell and tissue types, including mesenchymal, neuronal and epithelial cells. Human recombinant FGF10 protein drives the differentiation of embryonic stem cells (ESC) into gut-like structures, cardiomyocytes and hepatocytes [3]. It is also a potent factor in the development of organoids, increasing organoid size and branching phenotype compared to other FGFs [7].

[1] M. Igarashi, P. W. Finch & S. A. Aaronson. Characterization of recombinant human fibroblast growth factor (FGF)-10 reveals functional similarities with keratinocyte growth factor (FGF-7). J. Biol. Chem. 273, 13230–5 (1998). doi.org/10.1074/jbc.273.21.13230

[2] H-D. Beer, M. Bittner, G. Niklaus, C. Munding, N. Max, A. Goppelt & S. Werner. The fibroblast growth factor binding protein is a novel interaction partner of FGF-7, FGF-10 and FGF-22 and regulates FGF activity: implications for epithelial repair. Oncogene 24, 5269–77 (2005). doi.org/10.1038/sj.onc.1208560

[3] N. Itoh & H. Ohta. Fgf10: A Paracrine-Signaling Molecule in Development, Disease, and Regenerative Medicine. Curr. Mol. Med. 14, 504–509 (2014). doi: 10.2174/1566524014666140414204829

[4] X. Zhang, O. A. Ibrahimi, S. K. Olsen, H. Umemori, M. Mohammadi and D. M. Ornitz. Receptor specificity of the fibroblast growth factor family. The complete mammalian FGF family. J. Biol. Chem. 281, 15694–700 (2006). doi.org/10.1074/jbc.M601252200

[5] H. Ohuchi, T. Nakagawa, A. Yamamoto, A. Araga, T. Ohata, Y. Ishimaru, H. Yoshioka, T. Kuwana, T. Nohno, M. Yamasaki, N. Itoh and S. Noji. The mesenchymal factor, FGF10, initiates and maintains the outgrowth of the chick limb bud through interaction with FGF8, an apical ectodermal factor. Development 124, 2235–44 (1997). doi.org/10.1242/dev.124.11.2235

[6] H. Min, D. M. Danilenko, S. A. Scully, B. Bolon, B. D. Ring, J. E. Tarpley, M. DeRose, and W. S. Simonet. Fgf-10 is required for both limb and lung development and exhibits striking functional similarity to Drosophila branchless. Genes Dev. 12, 3156–61 (1998). doi: 10.1101/gad.12.20.3156

[7] A. Rabata, R. Fedr, K. Soucek, A. Hampl, Z. Koledova. 3D Cell Culture Models Demonstrate a Role for FGF and WNT Signaling in Regulation of Lung Epithelial Cell Fate and Morphogenesis. Front Cell Dev Biol. 2020 Jul 21;8:574. doi:10.3389/fcell.2020.00574.

Customer & collaborator data

FGF10 supports epithelial to mesenchymal transition (EMT) in human primary keratinocytes. Data and evaluation by Stemnovate Ltd.

EMT is a crucial morphogenetic process during development in which cells lose their epithelial characteristics and acquire migratory mesenchymal properties. Human FGF10 protein has an important role both during the embryonic EMT (type I) and on cancer cell initiation of metastasis (type III EMT).

human FGF10 induced EMT

Induction of EMT in human primary keratinocytes following treatment with hFGF10. Induction of EMT was evaluated using immunofluorescence staining to determine expression of the epithelial marker Cytokeratin 14 (CK14) and mesenchymal marker α-Smooth Muscle Actin (αSMA) in human primary epidermal keratinocytes after 4 days treatment with Qk003 hFGF10 (0-100 ng/ml).

FGF10 supports proliferation and promotes epithelial to mesenchymal transition in human primary keratinocytes. Data and evaluation by Stemnovate Ltd.

FGF10 induces proliferation of human primary keratinocytes

Cell proliferation assays to assess the effect of Qkine FGF10 (0-100 ng/ml) on human primary epidermal keratinocytes in serum-free keratinocyte media. Cells were evaluated at culture days: 0 (baseline), 1, 2, 3, 4 days, as summarized schematically in Figure 1a. Figure 1b shows cell proliferation for days 1, 2, 3, 4 and normalized to day 0 readouts (n=3; P*<0.05 vs control). The log concentration plot in Figure 1c shows percent cell proliferation normalized over untreated control (%) and to day 0 (baseline) after 4 days treatment (n=3; P*<0.05). The maximal cell proliferation was observed at ~10 ng/ml FGF10 and a reduction in cell number/viability as observed at 100 ng/ml. Data provided by Stemnovate Ltd, Cambridge, UK.

FGF10 bioactivity

Human FGF10 allowed us to derive and maintain patient-derived gastric organoids as part of our cocktail. Thanks to the high purity of their cytokines and quality-price ratio overcome by far our usual providers. Data from Dylan Liabeuf.

Additional resources

Publications

Human epidermis organotypic cultures, a reproducible system recapitulating the epidermis in vitro
In Experimental Dermatology on 28 April 2023 by Agarwal, R. et al.

FAQ

FGF10 is essential for normal embryonic development, particularly in limb bud development. FGF family growth factors have roles in cell survival and proliferation, angiogenesis, tumorigenesis, wound healing and tissue repair.

FGF10 is expressed in all branching morphogen organs such as the lungs, skin, ear and salivary glands.

FGF10 stimulates cell survival and proliferation on many cell types including lung and gut epithelium but has no effect on fibroblasts.

FGF-10 binds to and signals though FGF receptors 2b and 1b.

FGFRs phosphorylate specific tyrosine residues and activate the RAS-MAPK, PI3K-AKT, PLCγ, and STAT intracellular signaling pathways.

FGF10 drives the differentiation of embryonic stem cells into gut-like structures, cardiomyocytes and hepatocytes. It allows development of large organoids with branching phenotypes.

Our products are for research use only and not for diagnostic or therapeutic use.  Products are not for resale.

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