Staff Scientist Thermo Fisher Scientific Frederick, Maryland, United States
Abstract: Stem cells can self-renew and potentially differentiate into one or more mature cell lineages. The possibilities of stem cells, from basic research to clinical applications, require optimal culture conditions and the use of reagents, such as recombinant proteins. It is essential that these proteins act in a reliable and reproducible manner from the research phase all the way through to clinical applications. Basic fibroblast growth factor (bFGF or FGF-basic), also known as Fibroblast growth factor 2 (FGF2), plays a crucial role in stem cell biology, promoting survival, self-renewal, and pluripotency. Ensuring high-quality and consistency of bFGF is necessary to generate reliable results when working with a stem cell culture workflow; however, the uncertainty about the functional equivalence of different bFGF grades challenges scientists who transition from a research-grade to the clinical application grade. This study addresses this gap by assessing the effect of three Gibco™ PeproTech™ bFGF grades— RUO (Research Use Only; Recombinant Human FGF-basic), AOF (Animal free; Animal-Free Recombinant Human FGF-basic), and GMP (Good Manufacturing Practice; PeproGMP® Recombinant Human FGF-basic) on the growth, pluripotency, and differentiation potential of iPSCs reprogrammed from human dermal fibroblasts. iPSCs were first expanded over multiple passages in basal medium supplemented with one of three bFGF grades, during which growth rate and morphology were observed. Following expansion, iPSCs genetic stability, expression of pluripotency markers, and ability to differentiate into the three primary germ layers was measured. Our results showed that all grades of bFGF supported consistent iPSC growth, morphology, genetic stability, pluripotency, and differentiation potential, thus demonstrating that all three grades are functionally equivalent. Overall, the consistent performance of bFGF grades ensures the smooth transition from discovery to regenerative therapies, without the need for extensive revalidation. This improves operational efficiencies and supports cost-effective workflows, thereby paving the way for advancement of stem cell-based research from basic science to clinical applications.
