TITLE:
Instability of Fibroblast Growth Factor 2 in Cell Culture Media
AUTHORS:
A. S. Prakasha Gowda, Andrew D. Schaefer
KEYWORDS:
FGF-2 Stability, RP-HPLC, Cytotoxicity, Proliferation, Antioxidants
JOURNAL NAME:
Natural Science,
Vol.18 No.6,
June
16,
2026
ABSTRACT: Fibroblast growth factor 2 (FGF-2) is a key signaling molecule involved in wound healing, tissue remodeling, and the regulation of cell proliferation. Despite its important biological roles, the use of FGF-2 in cell culture and therapeutic settings is limited by its poor stability in aqueous environments. Three different antioxidants were selected and screened in this study for their ability to enhance the stability of FGF-2 in cell culture applications. BALB/3T3 and NIH/3T3 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented individually with disodium ethylenediaminetetraacetic acid dihydrate (EDTA), sodium selenite (Se), or zinc chloride (Zn), each at a final concentration of 0.1 μM. Cytotoxicity and cell proliferation were subsequently assessed using the MTT assay. The cells were then cultured at 37?C for three days to assess the biological effects of antioxidant supplementation. All three antioxidants individually enhanced the proliferation of both cell lines without inducing cytotoxicity. These findings suggest that antioxidants may represent a promising approach for improving vitro cell growth. Therefore, antioxidants were evaluated in parallel with FGF-2 by adding them separately to DMEM and Roswell Park Memorial Institute (RPMI) 1640 serum and cells free media under identical conditions to determine whether their proliferative effects were associated with improved FGF-2 stability. The stability of FGF-2 in each medium was assessed using reversed phase high performance liquid chromatography (RP-HPLC). The results show that, even in the presence of antioxidants, FGF-2 remained unstable at 37?C in both DMEM and RPMI media, indicating that these antioxidants do not confer stability to FGF-2 in either medium.