Stemfactor™ FGF-basic, Human Recombinant

ibroblast Growth Factor-basic (bFGF), also known as FGF-2, is a heparin-binding member of the FGF superfamily of molecules. Proteins of this family play a central role during prenatal development and postnatal growth and regeneration of a variety of tissues by promoting cellular proliferation and differentiation. Additionally, bFGF is a critical component of embryonic stem cell culture medium, allowing cells to remain in an undifferentiated state in serum-free medium^1,2. Stemfactor bFGF is an approximate 17 kDa protein consisting of 155 amino acid residues.

Stemgent and the Stemfactor brand name are trademarks of REPROCELL Inc., Japan.

Product Name: Stemfactor FGF-basic, Human Recombinant

Catalog Number: 03-0002

Size: 50 µg

Purity: Greater than 95% by SDS-PAGE analysis

Formulation: Lyophilized from 5 mM Tris-HCl, pH 7.5 with 150 mM NaCl.

Reconstitution: Centrifuge briefly and then reconstitute bFGF in 500 µL of 10 mM Tris, pH 7.6, to yield a stock solution of 0.1 mg/mL of bFGF. Avoid freeze-thaw cycles as it can result in loss of activity.

Storage and Stability: Stemfactor bFGF is shipped at room temperature. Lyophilized bFGF is stable for up to 6 months from date of receipt when stored at −20 °C to −80 °C. Reconstituted bFGF, at concentrations greater than or equal to 0.1 mg/mL, is stable for up to 3 months when stored at −20 °C and up to 6 months when stored at −80 °C.

Sterility: Tested to be negative for Mycoplasmasp. by PCR and microbial contamination by a sterility test.

Source: E. coli

Amino Acid Sequence: MAAGSITTLP ALPEDGGSGA FPPGHFKDPK RLYCKNGGFF LRIHPDGRVD GVREKSDPHI KLQLQAEERG VVSIKGVCAN RYLAMKEDGR LLASKCVTDE CFFFERLESN NYNTYRSRKY TSWYVALKRT GQYKLGSKTG PGQKAILFLP MSAKS

Uniprot Accession Number: P09038, residues 1-155

Endotoxin Level: Less than 1.0 EU/µg of bFGF as determined by the LAL method.

Biologic Activity: The ED50 is 0.1 to 1 ng/mL as determined by the dose dependent proliferation of NIH 3T3 cells.

Specification Sheets:

• 03-0002 Specifications Sheet

Safety Data Sheets:

• 03-0002 Safety Data Sheet

Application Notes:

• bFGF Supports Human ES Cell Self-Renewal

1. Amit, M., Carpenter, M.K., Inokuma, M.S., Chiu, C.P., Harris, C.P., Waknitz, M.A., Itskovitz-Eldor, J., and Thompson, J.A. Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. Dev Biol 227: 271-278 (2000).
2. Xu, C., Rosler, E., Jiang, J., Lebkowski, J.S., Gold, J.D., O'Sullivan, C., Delavan-Boorsma, K., Mok, M., Bronstein, A., and Carpenter, M.K. Basic fibroblast growth factor supports undifferentiated human embryonic stem cell growth without conditioned medium. Stem Cells 23: 315-323 (2005).
3. Vallier, L., Alexander, M., and Pedersen, R.A.  Activin/Nodal and FGF pathways corporate to maintain pluripotency of human embryonic stem cells. J Cell Sci 118: 4495-4509 (2005).
4. Wang, G., Zhang, H., Zhao, Y., Li, J., Cai, J., Wang, P., Meng, S., Feng, J., Miao, C., Ding, M., Li, D., and Deng, H. Noggin and bFGF cooperate to maintain the pluripotency of human embryonic stem cells in the absence of feeder layers. Biochem Biophys Res Commun 330: 934-942 (2005).
5. Greber, B., Lehrach, H., and Adjaye, J. Fibroblast growth factor 2 modulates transforming growth factor β signaling in mouse embryonic fibroblasts and human ESCs (hESCs) to support hESC self-renewal. Stem Cells 25: 455-464 (2007).

Additional Publications

• Prahl JD; Pierce Se; van der Schans EJC; Goetzee GA; Tyson T. The Parkinson's disease variant rs356182 regulates neuronal differentiation independently from alpha-synuclein. Human Molec Genetics ddac161:doi: 10.1093/hmg/ddac161 (2022).
• Li LK; Huang W-C; Hsueh Y-Y; Yanauchi Kk Olivares N; Davila R; Fang J; Ding X; Zhao W; Soto J; Hasani M; Novitch B; Li S. Intramuscular delivery of neural crest stem cell spheroids enhances neuromuscular regeneration after denervation injury. Stem Cell Research Therapy 13:205 (2022).
• Cheng C; Reis S; Adams ET; Fass DM; Angus SP ;Stuhlmiller TJ; Richardson J; Olafson H; Wang ET; Patnaik D; Beauchamp RL; Feldman DA; Silva MC; Sur M; Johnson GL; Ramesh V; Miller BL; Temple S; Kosik KS; Dickerson BC; Haggarty SJ. High-content image-based analysis and proteomic profiling identifies Tau phosphorylation inhibitors in a human iPSC-derived glutamatergic neuronal model of tauopathy. Sci Rep 11:17029 (2021).
• Pottmeier P Doszyn O; Peuckert C; Jazin E. Increased Expression of Y-Encoded Demethylases During Differentiation of Human Male Neural Stem Cells. Stem Cells Devel in Press://doi.org/10.1089/scd.2020.0138 (2020).
• McQuade A; Kang YJ; Hasselmann J; Jaioraman A; Sotelo A; Coburn M; Shabestari SK; Chadarevian JP; Fote G; Tu CH; Danhash E; Silva J; Martinez E; Gotman C; Prieto GA; Thompson LM; Steffan JS; Smith I; Davtyan H; Calahan M; Cho H; Blurton-Jones M. Gene expression and functional deficits underlie TREM2-knockout microglia responses in human models of Alzheimer’s disease. Natture Commun 11:5370 (2020).
• Bonnard C; Navaratnam V; Ghosh K; Chan PW; Tan TT; Pomp O; Ng AYG; Tohari S; Changede R; Carling D; Benkatesh B; Altunoglu U; Kayserili H; Reversade B. A loss-of-function NUAK2 mutation in humans causes anencephaly due to impaired Hippo-YAP signaling. J Exp Med 217:e20191561 (2020).
• Fukunaga I; Shirai K; Oe Y; Danzaki K; Ohta SZ; Shiga T; Chen C; Ikeda K; Akamatsu W; Kawano A; Kamiya K. Generation of two induced pluripotent stem cell lines from PBMCs of siblings carrying c.235delC mutation in the GJB2 gene associated with sensorineural hearing loss. Stem Cell Res 47:101910 (2020).
• Fukunaga I; Shiga T; Chen C; Oe Y; Danzaki K; Ohta S; Matsuoka R; Anzai R; Hibiya-Motegi R; Tajima S; Ikeda K; Akamatsu W; Kamiya K. Generation of the induced pluripotent stem cell (hiPSC) line (JUFMDOi004-A) from a patient with hearing loss carrying GJB2 (p.V37I) mutation. Stem Cell Res in press:doi.org/10.1016/j.xcr.2019.101674 (2019).
• Durens M; Nestor J; Herold K Niescier RF; Lunden FW; Phillips AW; Lin Y-C; Nestor MW. High-content interrogation of human induced pluripotent stem cell-derived cortical organoid platforms. bioRxiv http://dx.doi.org/10.1101/697623. : (2019).
• Schlick SF. Fibroblast-Cardiomyocyte Cross-Talk in Heart Muscle Formation and Function. Ph.D. Thesis, Georg-August University Göttingen :   (2018).
• Song Y; Subramanian K; Berberich MJ; Reoriguez S; Latorre IJ; Luria CM; Everly R; Albers MW; Mitchison TJ; Sorger PK. A dynamic view of the proteomic landscape during differentiation of ReNcell VM cells, an immortalized human neural progenitor line. Scientific Data 6:190016 (2019).
• Kilander MB; Wang C-H; Chang C-H; Nestor JE; Herold K; Tsai J-W; Nestor MW; Lin Y-C. A rare human CEP290 variant disrupts the molecular integrity of the primary cilium and impairs Sonic Hedgehog machinery. Sci Rep 8:17335 (2018).
• Yang G; Hong H; Trres A; Malloy KE; Choudhury CR; Kim J; Daadi MM. Standards for Deriving Nonhuman Primate-Induced Pluripotent Stem Cells, Neural Stem Cells and Dopaminergic Lineage. Int J Mol Sci 19:2788 (2018).
• Pierce SE; Tyson T; Booms A; Prahl J; Coetzee GA. Parkinson's disease genetic risk in a midbrain neuronal cell line. Neurobiol of Disease doi.org/10.1016/j.nbd.2018.02.007: (2018).
• Velasquez-Mao AJ; Tsao CJM; Monroe MN; Legras X; Bissig-Choisat B; Bissig K-D; Ruano R; Jacot JG. Differentiation of spontaneously contracting cardiomyocytes from non-virally reprogrammed human amniotic fluid stem cells. PLoS ONE 12(5):e177824 (2017).
• Noack C; Haupt LP; Zimmerman W-H; Streckfuss-Bomeke; Zelarayan LC. Generation of a KLF15 homozygous knockout human embryonic stem cell line using paired CRISPR/Cas9n, and human cardiomyocytes derivation. Stem Cell Research 23:127-131 (2017)
• Momcilovic O; Sivapatham R; Oron TR; Meyer M; Mooney S; Rao MA; Zeng X. Derivation, characterization, and neural differentiation of integration-free induced pluripotent stem cell lines from Parkinson's disease patients carrying SNCA, LRRK2, PARK2, and GBA mutations. PLoS ONE 11(5): e0154890. doi:10.1371/journal.pone.0154890 (2016)
• Bhutani K; Nazor KL; Williams R; Tran H; Dai H; Dzakula ZZ; Cho EH; Pang AWC; Roa M; Cao H; Schorck NJ; Loring JF. Whole-genome mutational burden analysis of three pluripotency induction methods. Nature Commun 7:10536 (2016)
• Zhu S; Russ HA; Wang X; Zhang M; Ma T; Xu T; Tang S; Hebrok M; Deng S. Human pancreatic β-like cells converted from fibroblasts. Nature Commun 7:10080 (2016)
• Chiang P-M; Wong PC. Differentiation of an embryonic stem cell to hemogenic endothelium by defined factors: Essential role of bone morphogenetic protein 4. Development 138: 2833-2843 (2011)
• Wang, Y-C., Nakagawa, M., Garitaonandia, I., Slavin, I., Altun, G., Lacharite, R. M., Nazor, K. L., Tran, H. T., Lynch, C., L., Leonardo, T. R., Liu, Y., Peterson, S. E., Laurent, L., C., Yamanaka, S., Loring, J. F. Specific lectin biomarkers for isolation of human pluripotent stem cells identified through array-based glycomic analysis. Cell Res 21(11): 1551 - 1563 (2011).