FGF-2 TOP®
(Thermostability Optimized )
FGF-2 TOP® is a novel, thermostabilized growth factor that allows you to grow FGF-2-dependent cell cultures more efficiently with fewer media changes.
Typical Applications
Pluripotent stem cells
Organoids
Neural stem cells
Uniquely Engineered for Stem Cell Production.
Through novel protein engineering, FGF-2 TOP® offers an increased half-life and therefore fewer feedings required compared to the wild-type growth factor.
It is significantly more tolerant of heat while maintaining full bioactivity, leading to improved homogeneity in your cell growth. Derived from our proprietary plant-based production system, FGF-2 TOP® is also fully animal component-free, endotoxin-free, and sustainably produced.
A Convenient Alternative with Cost Advantages
In order to maintain stem cell homogeneity and avoid spontaneous differentiation, scientists traditionally had to maintain a strict daily feeding schedule due to the short half-life (approximately 9 hours at 37°C) and temperature sensitivity of wild-type FGF-2.
Increased half-life
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To improve stability, a novel nine amino acid substitution of the wild-type FGF-2 was performed, improving the heat stability of FGF-2 TOP resulting in an increase in half-life under cell culture conditions to 37°C.
Save time and money
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By nature of its increased half-life and stability, FGF-2 TOP presents a constant exposure of growth factor (green line) in contrast to the short half-life and signaling of the wild-type protein that must be replenished daily (grey line). Thus, feeding schedules are more streamlined and cell culture phenotype is more homogenous.
The Stability to Support Your Development and Scale-Up
Retained superior bioactivity
Comparison of FGF-2 TOP®with a FGF-2 heat stable competitor and human FGF-2 wild-type in a 3T3 cell proliferation assay using varying concentrations of each protein after a 48-hour incubation at 37°C. FGF-2 TOP demonstrates maintenance of full bioactivity and with a 5-fold lower EC50, FGF-2 TOP® demonstrates a greater capacity to promote 3T3 cell proliferation and at lower concentrations than competing heat stable alternatives and wild-type FGF-2 in this model.
Supporting Proliferation of Adherent Cell Culture
In an internal MSC model analyzing confluence over 9 days, the addition of FGF-2 TOP® to an internal MSC formulation resulted in an equivalent cell confluence and expansion (as assessed through cell attachment) compared to the positive control containing 10% FBS. Similar results have been observed in both iPSC and adherent HEK-293 models (assessed but not shown).
Maintains Pluripotency and Stemness
FGF-2 TOP® maintains high-quality iPSC cultures under various concentrations and media feeding regimes as assessed by flow cytometry. Cells were treated with wild-type “human” FGF-2 or FGF-2 TOP® at 50 or 100 ng/mL either daily if not indicated or every other day (EOD). The negative control contained no FGF-2 supplementation. All tested conditions resulted in high expression of pluripotency markers. Most notably, EOD feeding with FGF-2 TOP® showed equivalent levels as well as provided further proof of bioactivity maintenance after stability changes. Source: App Note: Core Biogenesis in collaboration with TreeFrog Therapeutics.
Reduced costs and weekend-free feedings
Realize tangible benefits of FGF-2 TOP® protein stability: significant reductions in the amount of media required to feed cells, as well as reductions in the number of feedings, saving labor costs and obviating the need for inconvenient weekend feedings. Refer to the example feeding schedule and cost comparison of wild-type versus TOP®:
The Performance to Optimize Your PSC Cultures & Differentiation
Intensified iPSCs expansion
FGF-2 TOP® provides superior performance for high-quality iPSC cultures. A) When iPSCs are cultured in adherent conditions (2D) and supplemented with 100ng/mL of FGF-2 TOP®, cells display typical colony-forming morphologies, preventing the appearance of differentiated cells. B) When iPSCs are expanded in suspension (3D) and supplemented with 100ng/mL of FGF-2 TOP®, cells form embryoid bodies (EBs) with consistent shape and size.
Robust differentiation into NSC phenotypes
FGF-2 TOP® supports differentiation of iPSCs into Neural Stem Cells. E) Neural rosettes are formed from dissociated EBs when supplemented with 20ng/mL of FGF-2 TOP®. F) Cells transferred from the neural rosettes into pO/Lam-coated plates, start to differentiate into Neural Progenitor Cells (NPCs) reaching 80-90% of confluence after culture with 20ng/mL of FGF-2 TOP®.
The Consistency for Your Results while Working with MSCs Cultures
Increased cell numbers during MSC maintenance
Exogenous supplementation FGF-2 TOP® promotes higher increase of cell proliferation rates than “Wild-Type” FGF-2 on hMSCs. After 3 days in culture, hMSCs cultured with 10ng/mL of the wild-type version of hrFGF-2 (reference standard) promoted a cell expansion from 2x10^5 (seeding density) to 9.81x10^5. When hMSCs were supplemented with 10ng/mL of FGF-2 TOP® (Core Bio) cells expanded from the same seeding density to 18.42x10^5.
Recombinant FGF-2 TOP®
(Thermostability Optimized )
Available for Research Use + cGMP Use
FGF-2 TOP® is available in both Research Use Only and cGMP-compliant grades. To ensure seamless transition and scale-up capabilities, we have ensured that bioactivity and product performance of both grades are equivalent, as both grades are derived from the exact same molecule. The cGMP-compatible version is augmented with additional quality control testing and documentation to satisfy cell culture and manufacturing requirements in cGMP environments.
For cell therapy appliactions FGF-2 TOP® is manufactured under GMP conditions and governed by a robust quality management system in collaboration with Nucleus Bilogics.
Specifications
Type
Recombinant protein
Amino acid sequence
154 amino acids with 9 aa point mutations from the wild-type human FGF-2. Tag-free. Original reference sequence accession number: P09038
Origin
Plant-expressed; FGF-2 is produced in plant seeds of Camelina sativa.
Quality
RUO and GMP
Identity
Molecular weight. SDS-PAGE, HPLC available
Bioactivity
EC50 (<0.5 ng/mL)
Purity
>95-97%
Concentration
1 mg/mL
Mycoplasma
PCR
Bioburden
RUO: < 10 CFU/mL GMP: USP <61>
Endotoxin
RUO: < 100 EU/mL GMP: USP <85> ≤ 50 EU/mL
Shelf life
Stability studies ongoing for 24 months
Release
CoA
Storage
-20 to -80ºC
Highlighted applications
PSCs, MSCs, Organoids, HEKs
Featured studies
Evaluating the use of Thermostable FGF-2 (FGF-2 TOP®) for optimized iPSC Culture Protocols
In collaboration with TreeFrog Therapeutics
FGF-2 TOP® (Thermostability Optimized) enhances cell proliferation of MSCs and NHDFs
In collaboration withCreative Biolabs