FGF2 homologs in the context of cell culture
Sequence homologs of Basic Fibroblast Growth Factor (bFGF or FGF2) have exhibited differential performance in cell growth across species, with clear implications for improving cost and performance of human- and animal-derived cell culture. Continuing from work presented at ISSCR 2022 (see previous poster below), we evaluated several candidates for proliferation of human- and animal-derived cell lines. These results highlight the value of growth factor homolog screening when optimizing cell culture conditions. Presented at ISSCR 2023.
HiDef-B8 for single-cell clonal expansion
Fully defined and animal/serum-free media, manufactured under strict cGMP protocols and with traceable sourcing, are of high value in pluripotent stem cell isolation and expansion. Continuing from our collaboration with NanoCellect Biomedical (see application note below), we explored the value of our such medium, HiDef-B8, in the culture of hiPSCs. Presented at ISSCR 2023.
HiDef-B8 optimization using DOE
Consistency in growth media remains a challenge for stem cell research and manufacturing. Continuing from our work presented at ISSCR 2022 (see poster below), we continued our validation of HiDef-B8 using DOE-driven approaches, focusing on three components of the medium. Of the variants tested, HiDef-B8 continued to outperform based on function and cost. Presented at ISSCR 2023.
Gentle sorting of hiPSCs grown in HiDef-B8 medium
Gentle cell sorting technologies are advantageous in the isolation and expansion of homogeneous hiPSCs, as is the use of fully defined and animal-free cell media. In collaboration with NanoCellect Biomedical, we grew hiPSCs in HiDef-B8 and used the WOLF G2 to identify and enrich for hiPSCs expressing SSEA-4 and TRA-1-60-R. HiDef-B8 grown hiPSCs showed healthy morphology, viability, and expression of these markers, with robust post-sorting recovery in the same medium.
FGF2 variants for life science and cultivated meat
Naturally occurring isoforms of growth factors offer a range of unique features across diverse applications, and suggest means for evolution-driven improvements in stability, function, and efficacy. We produced a range of FGF2 homologs, coupled with the structure determination of thermostable human variant FGF2-G3, to inform the development of new growth factor solutions. Presented at ISSCR 2022.
Engineering approaches to medium optimization
Current growth medium solutions for the cell culture space are oftentimes inconsistent, variable, and costly, warranting the development of more optimized alternatives. We applied a design of experiments (DOE) to fit a response surface methodology (RSM) model to HiDef-B8 medium for continued optimization in stem cell culture. Presented at ISSCR 2022.
FGF2-G3 usage in HiDef-B8
Serum-containing media present major obstacles for the cell culture space, including batch-to-batch variability, undefined componentry, and the introduction of growth-inhibiting factors. We produced FGF2-G3, a thermostable variant of human FGF2, and studied its use in the context of HiDef-B8, a serum-free, fully-defined formulation for the culture of stem cells. Presented at ISSCR 2022.